Isotopically Enriched 64Zn-aspartate Attenuates Systemic Inflammation and Gut Dysbiosis in an Lps-induced Rat Model of Parkinson’s Disease

Isotopically Enriched 64Zn-aspartate Attenuates Systemic Inflammation and Gut Dysbiosis in an Lps-induced Rat Model of Parkinson’s Disease | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Isotopically Enriched 64 Zn-aspartate Attenuates Systemic Inflammation and Gut Dysbiosis in an Lps-induced Rat Model of Parkinson’s Disease Max Temnik, Mariia Rudyk, Alexandr Balakin, Sergey Gurin, Taisa Dovbynchuk, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8178695/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Mar, 2026 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract A growing body of research indicates that systemic inflammation contributes substantially to the progression of Parkinson’s disease (PD). Foundational studies propose that targeting inflammatory pathways may offer therapeutic benefits for PD and other neurodegenerative conditions. Our previous work demonstrated that a novel zinc aspartate compound enriched with the light isotope 64 Zn ( 64 Zn-asp) can counteract inflammatory and cognitive impairments triggered by intra-hippocampal Aβ 1−40 in rats, and can also mitigate neuroinflammation while promoting neuronal survival in a PD model. In the present study, we investigated the impact of this isotopically modified zinc compound on systemic inflammatory responses and gut microbiota composition in a rat model of PD induced by a single stereotactic intranigral injection of lipopolysaccharide (LPS). LPS-lesioned rats exhibited impaired locomotion, heightened anxiety-like behavior, and progressive dopaminergic dysfunction. 64 Zn-asp administration attenuated behavioral deficits and reduced apomorphine-induced rotations. Treatment normalized CRP levels, reversed LPS-induced increases in granulocytes and platelets, and corrected elevations in systemic inflammatory indices (including NLR, PLR, SII, and SIRI). 64 Zn-asp shifted circulating and peritoneal phagocytes toward an anti-inflammatory phenotype and partially restored thymus structure and cellularity. In the gut, LPS-induced PD resulted in marked reductions in Bifidobacterium and Lactobacillus spp. and an expansion of opportunistic Enterobacteriaceae and Staphylococcus spp. 64 Zn-asp largely preserved beneficial anaerobes and suppressed opportunistic taxa in both luminal and mucosa-associated compartments. These findings demonstrate that 64 Zn-aspartate exerts anti-inflammatory, immunomodulatory, and microbiota-stabilizing effects, suggesting potential therapeutic value as a disease-modifying strategy targeting neuroimmune-gut axis dysfunction in PD. Health sciences/Diseases Biological sciences/Immunology Biological sciences/Microbiology Health sciences/Neurology Biological sciences/Neuroscience Parkinson’s disease Inflammation Gut dysbiosis Stable light isotope-enriched zinc aspartate Motor function Anxiety-like behavior Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Parkinson’s disease (PD) is the second most common neurodegenerative disorder, affecting approximately 11.77 million people worldwide as of 2021 [ 1 , 2 ]. PD has seen the fastest growth in prevalence and associated disability among neurological disorders, emerging as a major global contributor to neurological impairment [ 3 ]. PD, once viewed primarily as a motoric disorder, is now understood to be a complex, multifactorial systemic condition. Its pathogenesis involves a range of interconnected mechanisms, including neuroinflammation, α-synuclein aggregation, dysfunction of the lysosomal-autophagy system, mitochondrial impairment with oxidative stress, and vesicular transport defects. An expanding body of research highlights the pivotal role of immune activation and inflammatory processes as key drivers in both the onset and progression of the disease. Persistently activated microglia release high levels of pro-inflammatory mediators that not only damage neurons but also perpetuate their own activation, establishing a self-amplifying cycle of neuroinflammation and neurodegeneration [ 4 ]. Activated microglia release diverse inflammatory mediators (matrix metalloproteinases, reactive oxygen and nitrogen species, chemokines, cytokines, etc.) and can phagocytose astrocytic end-feet, thereby compromising blood–brain barrier (BBB) integrity and increasing permeability [ 5 – 7 ]. In these circumstances, pro-inflammatory mediators released by chronically activated microglia may escape into the systemic circulation, initiating peripheral low-grade pro-inflammatory immune responses. In turn, chronic low-grade peripheral inflammation, especially under conditions of increased BBB permeability, can exacerbate neuroinflammation by enabling the circulating immune cells and inflammatory mediators to access the central nervous system. This bidirectional communication forms a vicious, self-perpetuating inflammatory loop that accelerates PD progression [ 8 , 9 ]. Current evidence-based treatment for PD focuses on symptomatic management with levodopa (L-DOPA) and does not specifically target pathological disease progression. However, long-term use frequently leads to complications such as dyskinesia, limiting its efficacy. Moreover, treatment usually begins in advanced stages, after substantial and irreversible neural damage has occurred. This underscores the urgent need for safer, more effective, and truly disease-modifying therapies [ 10 ]. One of the central therapeutic targets in the development of disease-modifying strategies for PD is the neuroimmune-inflammatory response. Inflammation plays a pivotal role in amplifying oxidative stress and creating a microenvironment conducive to neuronal injury. By modulating inflammatory pathways, it may be possible to attenuate oxidative modifications and disrupt the cascade of α-synuclein misfolding, aggregation, and intercellular propagation, thereby slowing or halting disease progression [ 11 ]. A range of immuno-inflammatory pathways is being explored as therapeutic targets in PD. Agents under investigation include the NLRP3 inflammasome inhibitor selnoflast [ 12 ], GLP-1 receptor agonists such as exenatide [ 13 ], the leukotriene receptor antagonist montelukast [ 14 ], PPARγ agonists [ 15 ], azathioprine [ 16 ], and selected traditional Chinese medicines [ 17 ], alongside trials of NSAIDs [ 18 ]. While some have shown modest benefits, most are limited by narrow target profiles and immunosuppressive side effects [ 19 ]. The gut-brain axis (GBA) has also emerged as a promising focus, with gut microbiota shown to modulate systemic immunity and microglial activity. Probiotics may relieve constipation, and certain strains exhibit neuroprotective potential, but robust clinical evidence for disease modification in PD is still lacking [ 20 ]. Collectively, these findings underscore the urgent need for effective anti-inflammatory therapies that can modify PD progression. In our previous work, we showed that intravenous administration of a novel zinc preparation – the isotopically-modified zinc aspartate enriched with 99.2% of the light zinc isotope 64 Zn (coded KLS-1) – reversed both inflammatory responses and cognitive impairments induced by intra-hippocampal Aβ 1−40 in rats [ 21 ], as well as alleviated neuroinflammation and motor dysfunction in rats with LPS-induced PD [ 22 ]. A meta-analysis revealed that serum zinc levels are significantly lower in PD patients than in healthy controls. Zinc, abundant in the hippocampus and cerebral cortex, is essential for behavior, learning, memory, and emotional regulation, and its deficiency may contribute to neurodegenerative disorders such as PD, Alzheimer’s disease, and amyotrophic lateral sclerosis [ 23 ]. In animal models of neurodegenerative diseases, zinc supplementation has shown neuroprotective and disease-modifying effects [ 24 , 25 ]. Zinc supplementation in elderly individuals significantly improves health outcomes by reducing the incidence of infectious diseases and mitigating inflammaging-associated pathologies, such as age-related macular degeneration, cardiovascular diseases, and others [ 26 , 27 ]. Zinc supplementation lowers serum inflammatory and oxidative stress markers in adults, suggesting a modulatory effect on systemic inflammation and oxidative pathways [ 28 , 29 ]. In a case report by Quiroga et al. (2014), resolution of movement disorder symptoms in a PD patient was reported following treatment with zinc sulfate in combination with vitamin C [ 30 ]. All these studies have used zinc with its natural isotopic composition, which is primarily an aggregate of heavy zinc isotopes. The brain normally favors the lighter 64 Zn isotope (the ratio of 66 Zn/ 64 Zn δ 66 Zn < 1) due to its isotope bonding preference with sulfur-containing amino acids like cysteine in metallothioneins [ 31 , 32 ]. With age and in neurodegenerative diseases, δ 66 Zn increases as heavier isotopes accumulate, likely due to altered binding preferences [ 33 , 34 ]. In Alzheimer’s disease, for example, heavy zinc isotopes preferentially bind to histidine residues in amyloid-β plaques [ 35 ]. Similar mechanisms may influence α-synuclein aggregation in PD, where histidine-50 is a key binding site for heavy zinc [ 36 – 38 ]. This study aimed to assess the effects of intravenous 64 Zn-asp (KLS-1) administration on systemic inflammation and gut microbiota in a rat model of PD induced by LPS. Materials and Methods Test Agent The therapeutic agent used in this study was isotopically modified 64 Zn di-aspartate ( 64 Zn-asp), an investigational zinc aspartate complex. The molecule consisted of one atom of 64 Zn chelated with two L-aspartic acid molecules (NeoFroxx, Einhausen, Germany), with the molecular formula C 8 H 12 O 8 N 2 64 Zn and a molar mass of 328 g/mol. Zinc accounted for 17.98% of the compound by weight, with the zinc component enriched to 99.2% 64 Zn. The compound, designated KLS-1, was synthesized by Pharmaceutical Factory Biopharma LLC (Bila Tserkva, Ukraine). Animals, Experimental Design, and LPS-Induced Parkinson’s Disease Model The study was conducted using adult male Wistar rats (8 weeks old, weighing 220–250 g) obtained from the vivarium of the Educational and Scientific Centre “Institute of Biology and Medicine” at Taras Shevchenko National University of Kyiv, Ukraine. Animals were housed under standard laboratory conditions with unrestricted access to food and water (standard chow). All animal experiments in this study were performed in accordance with the ARRIVE guidelines. A total of 51 rats were randomly assigned into four experimental groups using the “RAND()” function in Microsoft Excel (Fig. 1 ): Group I – intact controls (n = 12), did not underwent any manipulations; Group II – sham-operated controls (n = 12), intra-nigral H 2 Odd injection; Group III – LPS-induced PD model (n = 12), intra-nigral LPS injection (10 µg); Group IV – LPS-induced PD model treated with 64 Zn-asp (n = 15), intra-nigral LPS injection (10 µg) + 10 daily i.v. 64 Zn-asp injections (1.5 mg/kg). To induce unilateral lesions of the nigrostriatal pathway in groups III and IV, stereotaxic injections of 10 µg lipopolysaccharide (LPS, Escherichia coli O111:B4, Sigma) dissolved in 2 µL sterile saline (JSC “Infusion”, Ukraine) were administered directly into the substantia nigra as described previously [ 39 ]. Animals in group II received equivalent injections of sterile saline only. Prior to surgery, rats in groups II–IV were anesthetized with a combination of ketamine (75 mg/kg, Sigma, USA) and 2% xylazine (400 µL/kg, Alfasan International BV, Netherlands), and positioned in a stereotaxic apparatus (SEJ-4, Ukraine). Coordinates for injection were based on Hoban et al. (2013): AP − 5.3 mm, ML ± 2.0 mm from bregma, and DV − 7.2 mm below the dura [ 40 ]. The injection rate was 1 µL/min, with the needle left in place for 5 minutes post-injection to facilitate diffusion and prevent backflow. An 8-day period post-injection was allowed for disease development. Starting on Day 9, rats in group IV received daily intravenous injections of 64 Zn-asp (1.5 mg/kg) via the lateral tail vein for 10 consecutive days. Parkinsonian pathology was confirmed through behavioral assessments and post-mortem analysis of the nigrostriatal system using semi-quantitative tyrosine hydroxylase (TH) immunohistochemistry (data are not presented). On day 28, the rats were sacrificed using carbon dioxide inhalation and subsequent cervical dislocation [ 41 ], after which biological samples—including brain tissue, thymus, peritoneal lavage fluid, sections of the gastrointestinal tract, and blood—were obtained for analysis. Ethics statement All experimental procedures were approved by Ethics Committee of the Taras Shevchenko National University of Kyiv (protocol No. 4, 10.10.2021) and complied with the Animal Welfare Act, as well as national (Kyiv National Bioethics Congress, 2001–2007) and international (EU Directive 86/609/EEC) guidelines on the ethical treatment of laboratory animals. Behavioral Assessments Behavioral testing was conducted to evaluate locomotion, anxiety-like behavior, and dopaminergic dysfunction associated with the LPS-induced Parkinson’s disease model. Open Field Test On Day 24 post-surgery, spontaneous locomotor activity and anxiety-related behavior were assessed using the Open Field test, following the methodology adapted from [ 42 ]. The test arena consisted of a square enclosure (100 × 100 cm) with 30 cm high walls, illuminated by two overhead 60W LED lamps positioned 2 meters above the floor. A 6 × 6 grid (36 squares) was marked on the floor to aid spatial tracking. Each rat was individually placed in the center of the arena and allowed to explore for 5 minutes. Movements were recorded from above using a digital camera (Casio® EX-Z850, China) mounted 1 meter above the arena. Video data were later analyzed using MATLAB software. Measured parameters included total distance traveled, time spent in the central zone (inner perimeter), thigmotaxis (time spent near walls), number of rearings and grooming episodes, and frequency of defecation [ 43 , 44 ]. The arena was cleaned and dried between trials to eliminate olfactory cues. Apomorphine-Induced Rotation Test This test was used to evaluate the extent of dopaminergic neuron loss. It was conducted on Days 8 and 21 following stereotaxic surgery. Apomorphine hydrochloride (Sigma, USA) was administered intraperitoneally at a dose of 0.5 mg/kg. Five minutes after injection, each rat was placed in a cylindrical observation chamber (40 cm diameter), and contralateral (counterclockwise) rotations were recorded manually for 30 minutes using a stopwatch. Rats displaying more than 6 full-body turns per minute (rpm) were considered to have severe dopaminergic neuron (DN) loss (approximately 86.6%), while animals with ≤ 2 rpm were classified as having moderate DN loss (~ 44%) [ 45 ]. Elevated Plus Maze (EPM) Test Anxiety-like behavior was further assessed using the Elevated Plus Maze on Day 24 post-surgery, in accordance with Walf and Frye (2007) [ 46 ]. The maze consisted of a cross-shaped platform elevated 50 cm above the floor, featuring two open arms (50 × 10 cm) and two enclosed arms (50 × 10 × 30 cm), all connected by a central square (10 × 10 cm). Rats were placed in the center square facing an open arm at the start of the 5-minute test session. The setup was illuminated using two ceiling-mounted 60W LED lamps. Behavioral activity was recorded via IP camera and analyzed using MATLAB. Key parameters included total distance traveled, frequency of transitions between arms (open to closed and vice versa), time spent in open vs. closed arms, number of risk assessment behaviors (stretched attend postures), and total number of arm entries. All animals were given a short habituation period to minimize stress before testing. CRP Level Measurement C-reactive protein (CRP) levels in blood plasma, collected using the anticoagulant EDTA, were measured by enzyme-linked immunosorbent assay (ELISA) with the ELISA-CRP test system (Labcare Diagnostics India Pvt Ltd), following the manufacturer’s instructions. Lymphoid organ assessment Thymus and spleen tissues were carefully excised, weighed, and prepared for a single-cell suspension. Each thymus (spleen) was placed onto a 200-mesh sieve and gently homogenized using a tissue grinder in cold phosphate-buffered saline (PBS, pH 7.3) containing 2% heat-inactivated fetal calf serum (FCS; Gibco, Grand Island, NY, USA) until no visible clumps remained. The resulting single-cell suspensions were then counted using a standard hemocytometer. Cell viability, assessed via Trypan blue exclusion, consistently exceeded 95%. The measured parameters included the relative weights of the thymus and spleen, as well as the relative numbers of thymocytes and splenocytes. These values were calculated as follows: Relative organ weight = (absolute organ weight/animal weight) x 10 3 Relative cell count = (absolute cell count / absolute organ weight) x 10 − 10 Hematological Analysis Whole blood samples anticoagulated with EDTA were analyzed to assess hematological parameters. A fully automated hematology analyzer (Particle Counter Model PCE 210, ERMA, Japan), calibrated for rodent blood profiling, was used to evaluate blood cell indices in rats. Derived immune-inflammatory ratios were calculated using standard formulas based on absolute cell counts: Neutrophil-to-lymphocyte ratio (NLR) = absolute neutrophil count, ANC / absolute lymphocyte count, ALC Lymphocyte-to-monocyte ratio (LMR) = ALC / absolute monocyte count, AMC Platelet-to-lymphocyte ratio (PLR) = absolute platelet count, APC / ALC Platelet-to-neutrophil ratio (PNR) = APC / ANC Platelet-to-monocyte ratio (PMR) = APC / AMC Neutrophil-to-monocyte ratio (NMR) = ANC / AMC Systemic Immune-Inflammation Index (SII) = (ANC × APC) / ALC Systemic Inflammatory Response index (SIRI) = (ANC x AMC)/ ALC Neutrophil-platelet-to-lymphocyte–hemoglobin ratio (NPLHb) = (ANC × APC) / (ALC × Hb, g/dL) These indices were used to evaluate systemic inflammatory status and immune response patterns in the experimental groups. Peritoneal macrophage isolation Peritoneal macrophages (PMs) were harvested from the abdominal cavity of non-sensitized mice using a standard rapid peritoneal lavage technique, as previously described [ 47 ]. The peritoneal cavity was flushed with phosphate-buffered saline (PBS) supplemented with 100 U/mL heparin and 3% fetal bovine serum (FBS) to prevent clotting and preserve cell viability. The collected lavage fluid was centrifuged at 300 × g for 5 minutes at 4°C. The resulting cell pellet was washed twice with Hanks’ Balanced Salt Solution (HBSS) to remove residual serum and debris. Assessment of Phagocyte Metabolic Profile The functional status of phagocytes was evaluated by analyzing their phagocytic capacity, oxidative metabolism, and expression of surface phenotypic markers using flow cytometry. Phagocytosis was assessed based on a previously established method [ 47 ]. For this purpose, FITC-conjugated, heat-inactivated Staphylococcus aureus Cowan I (sourced from the microbiological collection of the Department of Microbiology and Immunology, ESC "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv) served as the phagocytic target. A suspension containing 2×10 5 microglia or macrophages was incubated with 5 µL of the bacterial stock (1×10 7 cells/mL) at 37°C for 30 minutes. Phagocytic activity was halted by adding a quenching solution composed of PBS with 0.02% EDTA and 0.04% paraformaldehyde. Phagocytic efficiency was quantified in two ways: the phagocytosis index (PhI), representing the average fluorescence intensity per phagocytic cell (reflecting the number of bacteria ingested), and the phagocytosis percentage (PP), denoting the proportion of fluorescently positive cells. Reactive oxygen species (ROS) generation, indicative of oxidative metabolic activity, was assessed using the fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA, Invitrogen) as previously described [ 47 ]. To determine the phenotypic characteristics of phagocytes, cells were stained with FITC-conjugated anti-CD86, PE-conjugated anti-CD80, and Alexa Fluor 647-conjugated anti-CD206 antibodies (BD Pharmingen, USA). All samples were analyzed using a FACSCalibur flow cytometer, and data were processed with CellQuest software (BD Biosciences, USA). Fecal water content assessment Fecal water content was determined using a modified method of Zhu et al. (2012) [ 48 ]. Each rat was placed individually in a clean cage lined with filter paper. Freshly expelled fecal pellets were collected immediately and placed in sealed tubes. The total sample was weighed to obtain the wet weight (WW), then dried at 60°C for 24 h and reweighed to determine the dry weight (DW). Stool water content (%) was calculated using the formula: SW = 100 − (DW × 100 / WW) Analysis of Luminal and Mucosa-Associated Gut Microbiota Microbiota analysis was performed using conventional culture-based techniques as described earlier [ 49 ]. To evaluate both luminal and wall-adherent microbial populations, distinct sections of the gastrointestinal tract were sampled. For mucosal microbiota assessment, colon segments measuring 1 cm 2 (located 2 cm from the anal verge) and sections of the small intestine (located 2 cm from the ileocecal valve) were excised, washed three times with chyme in saline, and homogenized using a Potter homogenizer. For luminal microbiota analysis, fecal samples were weighed and homogenized in 9 mL of sterile 0.5% sodium chloride solution to obtain a 10 − 1 dilution. Serial tenfold dilutions (ranging from 10 − 2 to 10 − 11 ) were prepared using the same procedure. Aliquots (10 µL) from each dilution were aseptically inoculated onto selective and differential culture media (HiMedia Laboratories Pvt. Ltd., India), including Bifidobacterium Agar, MRS Agar, Endo Agar, Mannitol Salt Agar, Iron Sulphite Agar, Simmons Citrate Agar, and Blood Agar Base (supplemented with 5% sterile defibrinated sheep blood). Cultures were incubated at 37°C for 24–48 hours. Microbial identification was conducted following the taxonomic keys provided in Bergey’s Manual of Determinative Bacteriology. Colony morphology, Gram staining, and a battery of biochemical tests were used for classification, including plasma coagulation, DNAse activity, lysozyme and phosphatase production, oxidase activity, carbohydrate fermentation profiles, Voges-Proskauer reaction, motility assessment, and novobiocin susceptibility (to differentiate S. aureus , S. epidermidis , and S. saprophyticus ). Lactose-negative E. coli strains were distinguished from other opportunistic Enterobacteriaceae by their ability to produce hydrogen sulfide. Results are expressed as the mean ± SD in logarithmic colony-forming units per gram of feces (lg CFU/g) and per square centimeter of intestinal tissue (lg CFU/cm 2 ). Statistical analysis Statistical analysis was performed using the Statistica 12.0 software. The Shapiro–Wilk test was applied to assess the normality of data distribution, following the approach described by Mishra et al. (2019) [ 50 ]. Variables with a normal distribution were expressed as mean ± standard deviation (SD), while those not normally distributed were reported as medians with interquartile ranges (IQR). For group comparisons, normally distributed data were analyzed using one-way ANOVA followed by Tukey’s post-hoc test for multiple comparisons. Non-normally distributed data were evaluated using the Mann–Whitney U test for pairwise comparisons and the Kruskal–Wallis test for comparisons across multiple groups, as outlined by Chan and Walmsley (1997) [ 51 ]. The Fisher’s Exact Test was employed to compare the proportions of rats showing an increase or decrease in rotation rate between the first and second apomorphine-induced rotation tests. A p-value of ≤ 0.05 was considered statistically significant. Results Administration of 64 Zn-aspartate attenuated dopaminergic system damage and improved behavioral outcomes in rats with LPS-induced Parkinson’s disease The apomorphine-induced rotation test is widely recognized as a standard method for evaluating dopaminergic system impairment and behavioral deficits in rat models of PD, including LPS-induced models [ 52 , 53 ]. On Day 8 post-surgery (prior to the initiation of treatment), rats administered LPS exhibited an average contralateral turning rate of 2.2 rpm, which corresponds to an estimated 44–60% loss of dopaminergic neurons (Table 1 ). By Day 15, 67% of LPS-lesioned rats demonstrated a 23% increase in rotation rate, suggesting continued neurodegeneration. The remaining 33% of animals in this group showed either stable or slightly reduced turning behavior. In contrast, among the rats receiving 64 Zn-asp, 86% showed a 13% decrease in rotation rate by Day 15, indicating potential neuroprotection or partial recovery of dopaminergic function. Only 14% of the treated animals displayed stable or slightly increased rotation rates, suggestive of ongoing neuronal loss. Table 1 Behavioral characteristics of rats with LPS-induced Parkinson’s disease treated with 64 Zn-asp Intact animals, n = 12 Sham-operated animals, n = 12 LPS-induced PD, n = 12 LPS-induced PD + 64 Zn-asp, n = 15 Open field test Total distance traveled, sm 3088.6 [2823.7; 3473.3] 2959.7 [2702.4; 4759.5] 2097.3 [1232.4; 2498.9] 2679.6 [1521.3; 3518.7] Time spent exploring the inner perimeter, sec 21.5 [9.3; 29.5] 14.0 [7.0; 37.0] 5.0 [0.5; 12.0] a 11.5 [7.3; 22.3] Time spent in squares surrounded by two walls, min 0.91 [0.90; 0.97] 0.95 [0.87; 0.96] 1.0 [0.98; 1.1] a b 0.98 [0.95; 1.0] Number of rearings 18.8 [16.5; 25.1] 21.3 [16.0; 28.3] 15.8 [10.3; 21.9] 16.2 [11.9; 23.3] Rearing duration, s 15.0 [12.5; 18.5] 11.8 [8.9; 18.8] 14.0 [13.9; 14.3] 14.8 [11.9; 16.8] Number of grooming episodes 4.0 [2.8; 6.3] 6.4 [4.4; 9.3] 8.8 [6.5; 9.3] a 6.2 [4.0; 7.3] Number of defecations 3.7 [1.8; 6.4] 4.5 [1.9; 6.7] 3.8 [1.8; 7.2] 5.2 [2.3; 8.4] Elevated Plus Maze (EPM) Test Total distance traveled, sm 1214.1 [1112.7; 1282.7] 1216.7 [957.6; 1256.0] 481.04 [401.3; 635.0] a b 999.7 [728.3; 1200.3] c Number of transitions 14.8 [10.8; 18.9] 16.6 [9.4; 21.6] 10.1 [6.2; 12.4] 13.6 [10.2; 19.9] Time in closed arms/time in open arms 5.1 [2.8; 6.1] 9.9 [5.8; 10.1] 15.2 [10.8; 18.3] a b 11.9 [7.8; 13.1] a Time spent in a stretched attend posture (risk assessment), s 200.3 [187.3; 326.1] 283.6 [211.2; 324.5] 61.6 [57.2; 115.1] a b 185.6 [135.9; 234.9] c Number of rearings 18.0 [17.0; 28.0] 25.0 [17.5; 30.5] 13.5 [9.3; 17.3] b 16.7 [10.3; 22.5] Apomorphine test Contralateral rotation rate, rpm Day 8 post lesion 1.23 [0.98; 1.65] 1.53 [1.23; 1.79] Day 21 post lesion 1.90 [1.28; 2.38] 0.85 [0.46; 1.23] c Percentage of rats exhibiting an increase/ decrease in rotation rate between the first and second apomorphine-induced rotation tests 67/33 14/86 c Note: data are presented as median and IQR or %. Data from different animal groups were compared using Kruskal-Wallis’s test or Fisher Exact Test for % correspondingly. a - p < 0.05 as compared to intact animals; b - p < 0.05 as compared to sham-operated animals; c - p < 0.05 as compared to untreated animals with LPS-induced PD. Gray shading in the cells indicates statistically significant differences. Motor function was assessed using the open field and EPM tests. LPS-induced dopaminergic damage was associated with impaired locomotion (Table 1 ). In the open field test, LPS-lesioned rats exhibited a 37% reduction in the median distance traveled compared to controls (p = 0.06). Similarly, in the EPM test, the distance traveled was reduced by approximately 2.4-fold relative to control animals (p ≤ 0.05). Treatment with 64 Zn-asp ameliorated these deficits: rats in the LPS-PD + 64 Zn-asp group demonstrated higher locomotor activity in both tests compared to untreated LPS-lesioned animals, with performance levels comparable to those of control groups. By the study’s end, LPS-lesioned rats exhibited moderate anxiety-like behavior. In the open field test, these animals spent about 2.5 times less time in the central area compared to controls (p ≤ 0.05), indicating increased anxiety (Table 1 ). The number of transitions between open and closed arms in the EPM was moderately reduced in LPS-lesioned rats. Conversely, LPS-lesioned rats treated with 64 Zn-asp tended to show increased transitions, suggesting reduced anxiety. Additionally, the ratio of time spent in closed arms to time in open arms was three times higher in LPS-lesioned rats than in controls, a pattern that was reversed with 64 Zn-asp treatment. Thigmotactic behavior, reflected by the time spent in corner zones (squares bordered by two walls), was slightly elevated in LPS-lesioned rats. Furthermore, these rats displayed reduced risk assessment behavior, as evidenced by a more than twofold reduction in the time spent in a stretched-attend posture compared to controls. Treatment with 64 Zn-asp normalized these parameters, bringing them in line with those observed in healthy rats. Rearing frequency, another indicator of exploratory behavior, was also reduced in LPS-lesioned animals. A tendency toward recovery of this behavior was observed in the treated group. Excessive grooming – commonly associated with anxiety [ 54 ] – was prominent in the LPS-PD group but normalized following 64 Zn-asp administration. Collectively, these findings suggest that 64 Zn-aspartate treatment not only protects against dopaminergic neurodegeneration but also alleviates motor impairments and mitigates anxiety-like behavior in the LPS-induced PD rat model. 64 Zn-Aspartate Treatment Improves Hematological Inflammatory Profiles in LPS-Induced Parkinsonian Rats The serum level of C-reactive protein (CRP), a widely recognized and reliable marker of systemic inflammation, exhibited substantial individual variability across all experimental groups (Fig. 2 ). Despite this variability, the median CRP level in the LPS-PD group was approximately 35% higher than that in control animals. In contrast, the median CRP level in the LPS-PD + 64 Zn-asp group was comparable to that of the control groups. White blood cell (WBC) counts and their differential components – neutrophils, lymphocytes, monocytes, and platelets – are well-established markers of systemic inflammation. In our study, LPS-lesioned rats exhibited a significant increase in both absolute and relative granulocyte (Gr) counts, a decrease in the relative lymphocyte (Ly) count, and a 1.7-fold increase in platelet (PLT) count (Table 2 ). These changes are indicative of a systemic inflammatory response. In contrast, LPS-lesioned animals treated with 64 Zn-aspartate showed no significant differences in these parameters compared to intact and sham-operated control groups, suggesting an anti-inflammatory effect of the treatment. Table 2 Hematological parameters in rats with LPS-induced Parkinson’s disease treated with 64 Zn-asp Intact animals, n = 12 Sham-operated animals, n = 12 LPS-induced PD, n = 12 LPS-induced PD + 64 Zn-asp, n = 15 WBC count with differentials WBC, x 10^3/µl 5.2 ± 1.6 5.9 ± 1.2 5.2 ± 1.2 7.0 ± 2.3 Ly, x 10^3/µl 3.9 ± 1.1 4.0 ± 0.7 3.3 ± 1.0 5.0 ± 1.0 c Mo, x 10^3/µl 0.4 ± 0.1 0.7 ± 0.2 a 0.4 ± 0.1 b 0.5 ± 0.2 Gr, x 10^3/µl 1.0 ± 0.2 1.2 ± 0.2 1.6 ± 0.2 a b 1.4 ± 0.2 c PLT, x 10^3/µl 140.3 ± 29.4 173.5 ± 60.0 240.6 ± 24.1 a b 160.6 ± 55.8 c Ly, % 73.9 ± 5.0 68.0 ± 4.4 60.5 ± 4.5 a b 70.2 ± 5.9 c Mo, % 8.0 ± 1.8 12.1 ± 1.7 a 7.7 ± 1.2 b 7.3 ± 1.9 b Gr, % 18.2 ± 4.6 20.0 ± 3.6 36.3 ± 5.4 a b 23.5 ± 5.0 c WBC-based indices of systemic inflammation NLR 0.25 [0.19; 0.29] 0.26 [0.25; 0.30] a 0.54 [0.41; 0.62] a b 0.34 [0.27; 0.39] a b c LMR 9.9 ± 2.8 5.9 ± 1.1 a 8.3 ± 2.2 b 10.3 ± 3.0 b PLR 37.7 ± 8.9 45.8 ± 11.9 51.8 ± 6.7 a 37.9 ± 11.3 PMR 347.8 ± 16.3 253.7 ± 34.8 614.5 ± 98.7 a b 534.3 ± 94.7 a b PNR 159.8 ± 47.3 158.6 ± 45.8 152.5 ± 67.3 116.4 ± 41.6 NMR 2.4 ± 0.7 1.8 ± 0.3 4.1 ± 0.8 a b 3.3 ± 0.7 b c SII 38.0 [18.3; 51.6] 39.1 [33.4; 48.6] 76.6 [75.9; 78.7] a b 49.0 [41.9; 57.9] a b c SIRI 0.11 [0.06; 0.15] 0.15 [0.13; 0.19] 0.22 [0.19; 0.23] a b 0.10 [0.07; 0.12] c NPLHbR 2.58 [1.97; 3.15] 3.46 [3.03; 4.10] 5.73 [5.29; 8.80] a b 3.24 [2.87; 3.53] c Note: data are presented as median and IQR or as mean ± SD. Data from different animal groups were compared using Kruskal-Wallis’s test or ANOVA with Tukey post-hoc test, respectively. a - p < 0.05 as compared to intact animals; b - p < 0.05 as compared to sham-operated animals; c - p < 0.05 as compared to untreated animals with LPS-induced PD. Ly – lymphocytes, Mo – monocytes, Gr – granulocytes, PLT – platelets. WBC-based indices are now recognized as valuable markers for assessing the severity of systemic inflammation in various conditions, including neurodegenerative diseases. Our study demonstrated a marked increase in nearly all calculated WBC-based inflammatory indices in LPS-PD rats (Table 2 ). In LPS-lesioned rats, NLR was increased 2.2-fold as compared to controls. The median PLR value was elevated by 27.2%. Compared to other inflammatory conditions, PMR and NMR are less commonly used in neurodegenerative disease research, yet they are considered informative markers of systemic inflammation. In our study, both PMR and NMR median values were approximately 2-fold elevated in LPS-lesioned rats. The median SII value was approximately 2-fold higher in the LPS-PD group compared to control animals, while another complex WBC-based index – SIRI – was 1.7 times higher than in controls. The median value of a newly proposed WBC-based inflammatory marker, NPLHbR, in the LPS-PD group was approximately 1.9 times higher than that in the control rats. Treatment with 64 Zn-asp effectively prevented LPS-induced alterations in WBC-based inflammatory indices, maintaining values at levels comparable to those of intact and sham-operated controls, unlike in untreated LPS-PD rats. 64 Zn-asp Modulates Polarization of Peripheral Blood Phagocytes in LPS-Induced PD Rats The functional state of circulating phagocytes was assessed using parameters commonly applied to characterize their polarized activation profile: phagocytic activity (PI), oxidative metabolism (ROS generation), and the expression of phenotypic markers CD80/86 and CD206. In the LPS-PD group, the median monocyte PI value was twice lower than that of control animals (Fig. 3 A). Treatment of LPS-lesioned rats with 64 Zn-asp increased the monocyte PI median by 31% compared to untreated parkinsonian rats, with values comparable to those in the sham-operated control group. The median neutrophil PI value in the LPS-PD group did not differ significantly from that of control rats, and treatment with 64 Zn-asp had no effect on this parameter (Fig. 3 B). Notably, neutrophil phagocytic activity was elevated in sham-operated animals, likely reflecting an N2 polarization shift associated with their participation in reparative processes following surgical intervention [ 55 ]. Monocyte ROS generation was markedly elevated in LPS-lesioned rats, indicating a pro-inflammatory shift characteristic of systemic inflammation (Fig. 3 C). Treatment with 64 Zn-asp substantially reduced oxidative metabolism: the median ROS generation in the LPS-PD + 64 Zn-asp group was sevenfold lower than in the LPS-PD group and approximately fourfold lower than in controls. Neutrophil ROS generation in LPS-PD rats was significantly higher than in controls. In contrast, LPS-lesioned animals treated with 64 Zn-asp showed ROS levels comparable to sham-operated rats (Fig. 3 D). Notably, sham-operated rats exhibited reduced ROS generation compared to intact animals, further supporting the notion of an anti-inflammatory, tissue-repairing metabolic profile. Neither the proportion of CD80/86⁺ cells (Fig. 3 E) nor the CD80/86 expression level (Fig. 3 F) in circulating phagocytes from LPS-lesioned rats differed significantly from controls. In the LPS-PD + 64 Zn-asp group, the median proportion of CD80/86⁺ cells was reduced by half compared with untreated animals, while the expression level was approximately threefold higher. The proportion of CD206⁺ circulating phagocytes was slightly elevated in LPS-lesioned rats, whereas treatment with 64 Zn-asp restored this value to control levels (Fig. 3 G). CD206 expression level in the LPS-PD group was similar to intact animals but 6.5-fold lower than in sham-operated rats (Fig. 3 H). Administration of 64 Zn-asp modestly increased CD206 expression. Since elevated CD206 is linked to an anti-inflammatory phagocyte phenotype, the higher values in sham-operated rats likely reflect post-surgical reparative processes, while the increase in the LPS-PD + 64 Zn-asp group may result from the drug’s anti-inflammatory action. Influence of 64 Zn-asp on Lymphoid Organ Weight and Cellularity PD-related inflammation was reflected in alterations of the lymphoid organs. LPS-PD rats showed significantly reduced thymus weight and increased thymus cellularity compared with controls (Table 3 ). Table 3 Lymphoid organ parameters in rats with LPS-induced Parkinson’s disease treated with 64 Zn-asp Intact animals, n = 12 Sham-operated animals, n = 12 LPS-induced PD, n = 12 LPS-induced PD + 64 Zn-asp, n = 15 Relative weight of thymus 1.32 [1.27; 1.36] 1.26 [1.18; 1.33] 1.07 [0.08; 1.08] a,b 1.16 [1.07; 1.29] c Relative number of thymocytes 1.75 [1.49; 2.01] 3.77 [1.72; 5.82] a 7.86 [6.13; 9.86] a,b 6.26 [4.24; 8.31] a,b Relative weight of spleen 2.97 [2.62; 3.33] 3.31 [3.07; 3.56] 2.55 [2.48; 2.59] b 3.19 [2.32; 3.45] c Relative number of splenocytes 3.68 [3.26; 4.10] 5.58 [5.53; 5.63] a 6.45 [5.69; 8.62] a,b 2.46 [2.44; 4.27] c Treatment with 64 Zn-asp restored thymus weight to control levels and partially normalized cellularity. Spleen weight was unchanged in LPS-lesioned rats compared to intact controls, but was marginally lower than in sham-operated animals; cellularity showed only a slight increase. In the LPS-PD + 64 Zn-asp group, both spleen weight and splenocyte counts were indistinguishable from controls. 64 Zn-asp Alters Polarization Profiles of Peritoneal Macrophages in LPS-Induced PD Rats Peritoneal macrophage PI was comparable across all experimental groups (Fig. 4 A). In LPS-PD rats, median ROS generation was approximately twice that of intact and sham-operated controls (Fig. 4 B), indicating heightened oxidative activity. Administration of 64 Zn-asp normalized ROS production to control levels. The proportion of CD206⁺ cells (representing large resident peritoneal macrophages) and CD206 expression levels in LPS-lesioned rats were similar to those in intact animals (Fig. 4 C, D). Treatment with 64 Zn-asp reduced the CD206⁺ cell fraction but increased CD206 expression. Neither the percentage of CD80/86⁺ cells (Fig. 4 E) nor CD80/86 expression levels (Fig. 4 F) in LPS-lesioned rats differed significantly from controls, and 64 Zn-asp treatment had no effect on these parameters. 64 Zn-Asp Mitigates Inflammation-Linked Gut Dysbiosis in a Rat with LPS-induced PD We analyzed the composition of the culturable gut microbiota in conjunction with measurements of fecal water content. At the end of the experiment, stool water content did not differ significantly among the experimental groups (Table 4 ). Nevertheless, rats in the LPS-PD group produced approximately 1.5-fold more feces (wet weight: 0.553 ± 0.11 g; dry weight: 0.268 ± 0.02 g) than controls (sham: 0.387 ± 0.18 g wet; 0.183 ± 0.05 g dry; p < 0.05), indicating impaired colonic motility, with stool accumulating in the colon rather than being efficiently propelled and expelled. In the LPS-PD + 64 Zn-asp group, fecal output was slightly lower than in untreated LPS-PD rats but remained higher than in controls. Table 4 Fecal wet weight, dry weight, and stool water content in rats with LPS-induced Parkinson’s disease treated with 64 Zn-asp Intact animals, n = 12 Sham-operated animals, n = 12 LPS-induced PD, n = 12 LPS-induced PD + 64 Zn-asp, n = 15 Fecal wet weight, g 0.356 ± 0.07 0.387 ± 0.18 0.554 ± 0.11 a, b 0.521 ± 0.08 a, b Fecal dry weight, g 0.191 ± 0.02 0.183 ± 0.05 0.268 ± 0.02 a, b 0.233 ± 0.02 Stool water content, % 54.1 ± 3.15 48.5 ± 7.47 48.9 ± 8.24 47.9 ± 6.57 Notes: Data are presented as mean ± SD. Data from different animal groups were compared using ANOVA with a Tukey post-hoc test. a - p ≤ 0.05 as compared to intact animals; b - p ≤ 0.05 as compared to sham-operated animals. In gut microbiota assessment, particular attention was focused on quantitative analysis of anaerobic saccharolytic genera Bifidobacterium and Lactobacillus , which synthesize γ-aminobutyric acid (GABA) involved in gastrointestinal motility and the gut-brain axis. In LPS-PD rats, counts of these bacteria were moderately reduced in the small-intestinal and large-intestinal wall-adherent microbiota (Fig. 5 A) and markedly decreased in the luminal microbiota of the large intestine (Fig. 5 B). Specifically, the number of Bifidobacterium species decreased by one order of magnitude — from lg 8.20 ± 0.6 CFU/g in sham-operated animals to lg 6.90 ± 0.20 CFU/g in the LPS-PD group. The number of Lactobacillus species decreased by two orders of magnitude compared to the control group — from lg 7.00 ± 0.40 CFU/g in controls to lg 5.50 ± 0.70 CFU/g in the LPS-lesioned rats. Treatment with 64 Zn-asp preserved bacterial abundance at control levels. Under normophysiological conditions, neither lactose-positive nor lactose-negative E. coli were detected in the mucosa-associated biotope (Fig. 5 C). In the LPS-PD group, lactose-positive E. coli reached 10 3 CFU/cm 2 , while lactose-negative strains increased to 10 5 CFU/cm 2 in the mucosa-associated microbiota of the small intestine, with trace amounts also detected in colonic tissue. LPS-PD was additionally associated with a pronounced increase in opportunistic enterobacteria, reaching lg 2.90 ± 1.00 CFU/cm 2 . In fecal microbiota from LPS-PD animals, total E. coli abundance did not differ significantly from sham-operated controls (Fig. 5 D). However, opportunistic enterobacteria counts increased by approximately two orders of magnitude, from lg 2.46 ± 0.73 CFU/g in controls to lg 4.30 ± 0.52 CFU/g in the LPS-PD group. Treatment with 64 Zn-asp resulted in a notable restoration of microbiota composition. Lactose-fermenting E. coli levels decreased, and lactose-negative strains were completely eliminated from the mucosa-associated microbiota. Moreover, 64 Zn-asp administration significantly reduced opportunistic enterobacteria abundance in both mucosa-associated and luminal compartments. Staphylococcus spp., including both mannitol-fermenting ( S. aureus ) and mannitol-negative strains, were also detected [ 56 ]. No Staphylococci were present in the mucosa-associated biotopes of control animals. In LPS-PD animals, Staphylococcus spp. were identified in the wall-adherent microbiota of the large intestine, suggesting increased aerobiosis (Fig. 5 E). Treatment with 64 Zn-asp slightly decreased the abundance of mannitol-fermenting staphylococci in the mucosa-associated biotope. Quantitative characteristics of staphylococci in the luminal microbiota did not differ significantly between groups (Fig. 5 F). Discussion LPS-induced PD models are among the most commonly used inflammatory models of PD, as they effectively reproduce key pathological features, including motor impairment, neuroinflammation, and systemic inflammation [ 39 , 57 ]. Furthermore, these models provide valuable tools for investigating anti-inflammatory and neuroprotective agents [ 58 ]. Given the growing recognition of systemic inflammation as a contributor to PD development and progression, as well as a potential therapeutic target, we employed this model to evaluate the effects of intravenous 64 Zn-asp administration on systemic immune-inflammatory responses in PD rats. Findings from in vitro and in vivo studies suggest that zinc may exert disease-modifying effects in PD through several mechanisms. Zinc appears to influence autophagy and lysosomal function, which may help limit α-synuclein accumulation, and it can reduce aggregation by enhancing albumin chaperone activity [ 59 , 60 ]. It has also been reported to modulate inflammatory pathways, including NF-κB signaling [ 61 ], the NLRP3 inflammasome [ 62 ], and STAT3 activation [ 63 ], while preserving immune competence [ 64 ]. In addition, zinc contributes to redox homeostasis by inducing metallothioneins and glutathione, thereby supporting antioxidant defenses [ 65 ]. In our study, intravenous administration of 64 Zn-asp exerted neuroprotective effects by reducing dopaminergic neurodegeneration, ameliorating motor dysfunction, and attenuating anxiety-like behavior in Parkinsonian rats. These outcomes were associated with the pronounced anti-inflammatory activity of the drug. Systemic inflammation in LPS-induced PD rats was confirmed by established markers, including elevated serum CRP levels, increased granulocyte and platelet counts, and a concomitant reduction in lymphocyte count [ 66 , 67 ]. Beyond these parameters, indices derived from complete blood counts provide a more nuanced assessment of immune-inflammatory status by capturing the relative proportions of different leukocyte populations. Among them, the NLR, coupled with relative lymphopenia, has been linked to neurodegeneration-associated protein alterations, particularly within α-synuclein and amyloid-β pathways, and correlates with greater clinical burden in PD patients [ 68 , 69 ]. The PLR represents another established indicator of peripheral immune dysregulation and systemic inflammation in PD [ 70 ]. Similarly, an elevated NMR suggests an intensified inflammatory response [ 71 ]. Platelets, in addition to their roles in hemostasis and thrombosis, are increasingly recognized as active mediators of inflammation, supporting the recruitment of lymphocytes, neutrophils, and monocytes to inflamed tissues, thereby amplifying immune responses. In PD, chronic inflammation promotes platelet hyperreactivity, which may further exacerbate neuroinflammation [ 72 ]. This highlights the relevance of the PMR as an informative marker of systemic inflammatory activity. Composite indices have also been proposed to better capture the complexity of immune-inflammatory dynamics. The SII, which incorporates neutrophil, platelet, and lymphocyte counts, provides an integrated measure of the immune-inflammatory balance. Elevated SII is strongly associated with increased PD risk, particularly in females [ 73 ]. Likewise, the SIRI, which combines neutrophil, monocyte, and lymphocyte counts, reflects the interplay between immune activation and suppression and may indicate states of immunodeficiency or immune exhaustion [ 74 ]. More recently, the NPLHbR has been introduced as a reliable WBC-based marker of systemic inflammation. Unlike other indices, NPLHbR also incorporates hemoglobin, a parameter frequently reduced in chronic inflammatory states, including age-related “inflammaging,” which is associated with anemia and sustained immune activation [ 75 ]. In our study, the median values of all WBC-based inflammatory indices were significantly (~ 2 times) elevated in LPS-lesioned rats, reflecting pronounced and persistent systemic inflammation accompanied by features of immune exhaustion. 64 Zn-aspartate prevented LPS-induced changes in WBC-based inflammatory indices, preserving values comparable to controls. In addition to the well-established anti-inflammatory properties of zinc mediated through modulation of key pro-inflammatory signaling pathways such as NF-κB and IL-6, promotion of anti-inflammatory protein expression, and regulation of nitric oxide production [ 65 ], one potential mechanism underlying the observed effects of 64 Zn-asp may involve its impact on thymic function. In the context of systemic inflammation, acute thymic atrophy, as observed in LPS-lesioned rats in our study and in the MPTP model of PD in mice [ 76 ], may occur, leading to extensive loss of developing T cells during thymic selection, along with infiltration of immune cells that can further disrupt thymic architecture [ 77 ]. Zinc plays a pivotal role in normal T-cell development and in recovery following acute injury and inflammation, as it promotes thymic regeneration by inducing endothelial cell production of bone morphogenetic protein 4 (BMP4) [ 78 ]. In addition, zinc can promote differentiation of regulatory T-cells [ 79 ], which interfere with inflammatory signaling pathways controlling systemic inflammation [ 80 ]. In addition to restoring immune cell counts in LPS-PD animals, intravenous administration of 64 Zn-asp abrogated the pro-inflammatory metabolic shift of phagocytic cells – key mediators of inflammatory responses – in both peripheral blood and the peritoneal cavity. In PD, systemic inflammation is driven in part by monocytes and neutrophils. Clinical studies consistently report elevated circulating levels of these cells along with increased concentrations of inflammatory markers. Among monocytes, the classical pro-inflammatory (M1) subset is particularly implicated, as these cells release cytokines and chemokines that exacerbate neuroinflammation and contribute to neuronal damage. Neutrophils, the most abundant immune cells in circulation, act as early responders to inflammatory stimuli. In PD, pro-inflammatory N1 neutrophils are believed to play a role in the initial immune response to potential triggers, including pathogens or cellular damage. Although their role in PD has been less extensively investigated than that of monocytes and microglia, they are increasingly recognized as important contributors to early inflammatory events [ 81 ]. In our study, LPS-lesioned rats exhibited an increased proportion of circulating CD206⁺ phagocytic cells, likely representing a subset of activated myeloid cells with a pro-inflammatory metabolic profile. As described by Trombetta et al. (2018), these cells display a mixed phenotype, co-expressing markers of both M1 (pro-inflammatory) and M2 (anti-inflammatory) phagocytes. A further indication of their pro-inflammatory metabolic bias is their reduced maturation relative to counterparts expressing the same M2 markers but displaying a more anti-inflammatory phenotype, as well as the absence of CD204 expression [ 82 ]. In addition, reduced phagocytic activity in monocytes, together with markedly elevated ROS production in both neutrophils and monocytes of LPS-lesioned rats, further supports the presence of a pro-inflammatory metabolic shift in these cells [ 83 , 84 ]. Treatment with 64 Zn-asp returned CD206 + phagocyte cell fraction to the norm, marginally increased monocyte phagocytic activity, and substantially decreased the oxidative metabolism of both phagocytic cell populations, which indicates an anti-inflammatory metabolic shift. In treated parkinsonian rats, we observed a reduced fraction of CD80/86⁺ cells accompanied by increased CD80/86 expression levels. This paradoxical pattern may indicate a shift toward the differentiation of CD80/86⁺ monocyte-derived myeloid suppressor cells, potentially driven by the drug’s anti-inflammatory action. Costimulatory molecules of the B7.1 family, CD80 and CD86, serve a dual role as markers of phagocyte polarization. On the one hand, upregulation of CD80/CD86 expression reflects the acquisition of antigen-presenting capacity by circulating phagocytes and is indicative of a pro-inflammatory metabolic profile [ 85 ]. On the other hand, the majority of ex vivo -generated myeloid-derived suppressor cells (MDSCs) exhibit high CD80/CD86 expression, and monocytic MDSCs in vivo are characterized by strong CD86 positivity [ 86 , 87 ]. The metabolic profile of peritoneal macrophages in LPS-lesioned rats was moderately altered compared with controls, with increased ROS production indicating a pro-inflammatory metabolic shift [ 88 ]. Treatment with 64 Zn-aspartate reduced ROS generation and enhanced CD206 expression, a reliable marker of an anti-inflammatory phenotype in tissue-resident macrophages [ 89 ]. The hypothesis that PD may originate in the periphery and involve environmental risk factors has brought increasing attention to the role of the gut microbiota. Intestinal dysbiosis, which can precede the clinical diagnosis of PD by several years, has been proposed as a potential trigger of the disease through the disruption of microbial and mucosal immune homeostasis, thereby initiating an inflammatory cascade that extends to the brain and contributes to the pathophysiology of PD [ 90 ]. Nevertheless, the prevailing view is that dysbiosis is more likely a consequence of PD rather than its primary cause. Although it may arise years before clinical onset and contribute to disease progression, it is not generally considered the initiating event [ 91 ]. Instead, inflammation inherent to PD appears to be a major driver of gut dysbiosis. Under inflammatory conditions, the relative abundance of obligate anaerobes belonging to the Bacteroidetes and Firmicutes phyla declines, while Proteobacteria – particularly Gammaproteobacteria such as Enterobacteriaceae – expand and become dominant within the gut microbiota [ 92 ]. In our study, systemic inflammation in parkinsonian rats was closely associated with pronounced intestinal dysbiosis, suggesting a bidirectional interplay between peripheral immune activation and gut microbial imbalance. Representatives of the anaerobic saccharolytic genera Bifidobacterium and Lactobacillus exhibited a marked reduction in both mucosa-associated and luminal microbiota. In parallel, members of the family Enterobacteriaceae demonstrated a significant expansion. Specifically, both lactose-fermenting and non-fermenting Escherichia coli , as well as opportunistic enterobacteria, were detected in the wall-adherent microbiota of LPS-lesioned animals, whereas these taxa were absent in control rats. A comparable increase in opportunistic enterobacteria was also observed in the colonic luminal microbiota. Both systemic and intestinal inflammation are known to alter the physiological oxygen gradient, leading to localized increases in oxygen availability. This shift generates conditions that are more aerobic than those observed in the healthy gut. Within this altered niche, members of the Enterobacteriaceae family gain a selective advantage, while commensal symbiotic bacteria are negatively affected by inflammation-induced environmental changes [ 93 ]. Among the pathobionts characteristic of the gut microbiota in patients with PD, adherent-invasive E. coli (AIEC) has been identified. AIEC is thought to contribute to PD through the production of substances such as curli, which promote the aggregation of α-synuclein, a key pathogenic process in the disease [ 94 ]. Inflammation in the peritoneal cavity, reflected by the pro-inflammatory metabolic profile of peritoneal macrophages and accompanied by pronounced dysbiosis, suggests concurrent inflammation within the intestinal mucosa-associated lymphoid tissue, which may promote the growth and virulence of AIEC. Notably, the emergence of Staphylococcus , atypical for mucosa-associated intestinal microbiota, likely reflects inflammation-driven aerobization of the biotope, further highlighting microbial community disruption. In LPS-lesioned rats, gut microbiota imbalance was associated with increased fecal output without a corresponding rise in water content, indicating gastrointestinal dysfunction – one of the most common non-motor symptoms in PD [ 95 , 96 ]. This dysbiosis may also contribute to the thymic atrophy observed in these animals [ 97 ]. Treatment of LPS-PD rats with 64 Zn-asp promoted the rebalancing of gut microbial communities, evidenced by an increase in bifidobacteria and lactobacilli in the luminal microbiota and a concomitant reduction or disappearance of enterobacteria in the wall-adherent compartment, suggesting a microbiota-stabilizing effect. This effect may result from the anti-inflammatory action of the compound, although a direct impact of the preparation on the microorganisms cannot be excluded. Zinc has been reported to exert bidirectional effects on Lactobacillus species, promoting the growth of some while inhibiting others [ 98 ]. In addition, zinc can suppress the growth and virulence of AIEC by reducing bacterial adherence, biofilm formation, and the expression of key virulence factors [ 99 ]. This study is subject to several limitations. First, it did not directly investigate how 64Zn-asp influences zinc homeostasis or neuronal function, leaving important aspects of its underlying mechanism unresolved and emphasizing the need for further exploration. Second, evaluating the metabolic profile of phagocytes requires incorporating additional phenotypic markers to enable more precise differentiation of myeloid-derived suppressor cells. Finally, molecular approaches are necessary to gain deeper insight into the impact of 64 Zn-asp on the intestinal microbiota. Conclusion Systemic inflammation is a recognized contributor to the onset and progression of PD, making it an important therapeutic target. Peripheral inflammatory events can intensify neuroinflammation and accelerate neurodegeneration; therefore, anti-inflammatory interventions are being actively explored. In this study, we demonstrated that a zinc preparation with a unique isotopic composition 64 Zn-asp – endowing it with high biological activity and potent anti-inflammatory properties – was able to attenuate systemic inflammation in animals with LPS-PD while also modulating gut dysbiosis. These findings highlight the potential of this preparation as a multifaceted therapeutic strategy; however, further mechanistic studies and translational research are essential to validate its efficacy and safety in the context of human PD. Declarations Data availability The data that support the findings of this study are available from the corresponding author, upon reasonable request. Author contributions MT: supervision and review. MR: methodology, investigation, formal analysis. AB: methodology, conceptualization, and project administration. SG: conceptualization, writing, review, and editing. TD: methodology, investigation. RB: methodology, conceptualization. ND: methodology, investigation. RD: methodology, investigation. TS: methodology, investigation, and writing original draft. GT: supervision and writing – review and editing. LS: methodology, conceptualization, project administration, and writing the original draft. All authors read the final version of the manuscript. Funding The study was supported by a TSNUK project N 18DP036-10. Competing interests The authors declare no competing interests. References Su, D. et al. 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1","display":"","copyAsset":false,"role":"figure","size":4171571,"visible":true,"origin":"","legend":"\u003cp\u003eAnimal groups and study design.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/13486bc25c440a1c7a1f2da9.png"},{"id":98432690,"identity":"cb51be18-32ab-4da3-b259-a2d3fd2c7813","added_by":"auto","created_at":"2025-12-17 16:49:48","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":172551,"visible":true,"origin":"","legend":"\u003cp\u003ePlasma CRP levels in rats with LPS-induced Parkinson’s disease treated with 64Zn-asp. Data are presented as medians and IQR. Data from intact, sham-operated, and LPS-lesioned animals were compared using Kruskal-Wallis’s test.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/f679513036afd4b03fe43e53.png"},{"id":98265174,"identity":"e5599770-0ec2-4acb-a066-c19d20f71fc9","added_by":"auto","created_at":"2025-12-15 22:32:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":3116829,"visible":true,"origin":"","legend":"\u003cp\u003eMetabolic characteristics of peripheral blood phagocytes in rats with LPS-induced Parkinson’s disease treated with 64Zn-asp. A – monocyte phagocytosis index; B – neutrophil phagocytosis index; C – monocyte ROS generation; D – neutrophil ROS generation; E – fraction of CD80/86-positive cells; F – CD80/86 expression level; G – fraction of CD206-positive cells; H – CD206 expression level. Data are presented as medians and IQR. a - p ≤ 0.05 as compared to intact animals, b - p ≤ 0.05 as compared to sham-operated animals; c - p ≤ 0.05 as compared to LPS-PD group (Kruskal-Wallis’s test).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/f03fc7f29068fd10d06a677b.png"},{"id":98265172,"identity":"a788e1d0-e869-4fff-8ad2-972373ba84f7","added_by":"auto","created_at":"2025-12-15 22:32:34","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2199948,"visible":true,"origin":"","legend":"\u003cp\u003eMetabolic characteristics of peritoneal macrophages in rats with LPS-induced Parkinson’s disease treated with 64Zn-asp. A – phagocytosis index; B – ROS generation; C - fraction of CD206-positive cells; D CD206 expression level; E – fraction of CD80/86-positive cells; F – CD80/86 expression level. Data are presented as medians and IQR. a - p ≤ 0.05 as compared to intact animals, b - p ≤ 0.05 as compared to sham-operated animals; c - p ≤ 0.05 as compared to LPS-PD group (Kruskal-Wallis’s test).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/9a3af9253d7b66ecf2ded338.png"},{"id":98432742,"identity":"c4234f94-50e6-4db2-b6e9-37ec8c204236","added_by":"auto","created_at":"2025-12-17 16:49:54","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1907059,"visible":true,"origin":"","legend":"\u003cp\u003eCulturable wall-adherent and luminal gut microbiota in rats with LPS-induced Parkinson’s disease treated with 64Zn-asp. A – wall-adherent Bifidobacterium and Lactobacillus; B – luminal Bifidobacterium and Lactobacillus; C - wall-adherent enterobacteria; D – luminal enterobacteria; E - wall-adherent staphylococci; F – luminal staphylococci. Data are presented as mean ± SD. Data from different animal groups were compared using ANOVA with a Tukey post-hoc test. a - p≤0.05 as compared to intact animals; b - p≤0.05 as compared to sham-operated animals; c - p≤0.05 as compared to untreated animals with LPS-induced PD.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/e0e8b6fae049cccb4994b6cf.png"},{"id":105755021,"identity":"050e3ee7-934f-436f-a26f-228cc59a8c97","added_by":"auto","created_at":"2026-03-30 16:24:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":10595945,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8178695/v1/7f48e86c-2f8a-4242-a812-bbc4cec02d21.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eIsotopically Enriched \u003csup\u003e64\u003c/sup\u003eZn-aspartate Attenuates Systemic Inflammation and Gut Dysbiosis in an Lps-induced Rat Model of Parkinson’s Disease\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eParkinson\u0026rsquo;s disease (PD) is the second most common neurodegenerative disorder, affecting approximately 11.77\u0026nbsp;million people worldwide as of 2021 [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. PD has seen the fastest growth in prevalence and associated disability among neurological disorders, emerging as a major global contributor to neurological impairment [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. PD, once viewed primarily as a motoric disorder, is now understood to be a complex, multifactorial systemic condition. Its pathogenesis involves a range of interconnected mechanisms, including neuroinflammation, α-synuclein aggregation, dysfunction of the lysosomal-autophagy system, mitochondrial impairment with oxidative stress, and vesicular transport defects. An expanding body of research highlights the pivotal role of immune activation and inflammatory processes as key drivers in both the onset and progression of the disease. Persistently activated microglia release high levels of pro-inflammatory mediators that not only damage neurons but also perpetuate their own activation, establishing a self-amplifying cycle of neuroinflammation and neurodegeneration [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Activated microglia release diverse inflammatory mediators (matrix metalloproteinases, reactive oxygen and nitrogen species, chemokines, cytokines, etc.) and can phagocytose astrocytic end-feet, thereby compromising blood\u0026ndash;brain barrier (BBB) integrity and increasing permeability [\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. In these circumstances, pro-inflammatory mediators released by chronically activated microglia may escape into the systemic circulation, initiating peripheral low-grade pro-inflammatory immune responses. In turn, chronic low-grade peripheral inflammation, especially under conditions of increased BBB permeability, can exacerbate neuroinflammation by enabling the circulating immune cells and inflammatory mediators to access the central nervous system. This bidirectional communication forms a vicious, self-perpetuating inflammatory loop that accelerates PD progression [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eCurrent evidence-based treatment for PD focuses on symptomatic management with levodopa (L-DOPA) and does not specifically target pathological disease progression. However, long-term use frequently leads to complications such as dyskinesia, limiting its efficacy. Moreover, treatment usually begins in advanced stages, after substantial and irreversible neural damage has occurred. This underscores the urgent need for safer, more effective, and truly disease-modifying therapies [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. One of the central therapeutic targets in the development of disease-modifying strategies for PD is the neuroimmune-inflammatory response. Inflammation plays a pivotal role in amplifying oxidative stress and creating a microenvironment conducive to neuronal injury. By modulating inflammatory pathways, it may be possible to attenuate oxidative modifications and disrupt the cascade of α-synuclein misfolding, aggregation, and intercellular propagation, thereby slowing or halting disease progression [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. A range of immuno-inflammatory pathways is being explored as therapeutic targets in PD. Agents under investigation include the NLRP3 inflammasome inhibitor selnoflast [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], GLP-1 receptor agonists such as exenatide [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], the leukotriene receptor antagonist montelukast [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], PPARγ agonists [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], azathioprine [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], and selected traditional Chinese medicines [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], alongside trials of NSAIDs [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. While some have shown modest benefits, most are limited by narrow target profiles and immunosuppressive side effects [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The gut-brain axis (GBA) has also emerged as a promising focus, with gut microbiota shown to modulate systemic immunity and microglial activity. Probiotics may relieve constipation, and certain strains exhibit neuroprotective potential, but robust clinical evidence for disease modification in PD is still lacking [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Collectively, these findings underscore the urgent need for effective anti-inflammatory therapies that can modify PD progression.\u003c/p\u003e\u003cp\u003eIn our previous work, we showed that intravenous administration of a novel zinc preparation \u0026ndash; the isotopically-modified zinc aspartate enriched with 99.2% of the light zinc isotope \u003csup\u003e64\u003c/sup\u003eZn (coded KLS-1) \u0026ndash; reversed both inflammatory responses and cognitive impairments induced by intra-hippocampal Aβ\u003csub\u003e1\u0026minus;40\u003c/sub\u003e in rats [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], as well as alleviated neuroinflammation and motor dysfunction in rats with LPS-induced PD [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. A meta-analysis revealed that serum zinc levels are significantly lower in PD patients than in healthy controls. Zinc, abundant in the hippocampus and cerebral cortex, is essential for behavior, learning, memory, and emotional regulation, and its deficiency may contribute to neurodegenerative disorders such as PD, Alzheimer\u0026rsquo;s disease, and amyotrophic lateral sclerosis [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. In animal models of neurodegenerative diseases, zinc supplementation has shown neuroprotective and disease-modifying effects [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Zinc supplementation in elderly individuals significantly improves health outcomes by reducing the incidence of infectious diseases and mitigating inflammaging-associated pathologies, such as age-related macular degeneration, cardiovascular diseases, and others [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Zinc supplementation lowers serum inflammatory and oxidative stress markers in adults, suggesting a modulatory effect on systemic inflammation and oxidative pathways [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. In a case report by Quiroga et al. (2014), resolution of movement disorder symptoms in a PD patient was reported following treatment with zinc sulfate in combination with vitamin C [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. All these studies have used zinc with its natural isotopic composition, which is primarily an aggregate of heavy zinc isotopes. The brain normally favors the lighter \u003csup\u003e64\u003c/sup\u003eZn isotope (the ratio of \u003csup\u003e66\u003c/sup\u003eZn/\u003csup\u003e64\u003c/sup\u003eZn δ\u003csup\u003e66\u003c/sup\u003eZn \u0026lt; 1) due to its isotope bonding preference with sulfur-containing amino acids like cysteine in metallothioneins [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. With age and in neurodegenerative diseases, δ\u003csup\u003e66\u003c/sup\u003eZn increases as heavier isotopes accumulate, likely due to altered binding preferences [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In Alzheimer\u0026rsquo;s disease, for example, heavy zinc isotopes preferentially bind to histidine residues in amyloid-β plaques [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Similar mechanisms may influence α-synuclein aggregation in PD, where histidine-50 is a key binding site for heavy zinc [\u003cspan additionalcitationids=\"CR37\" citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThis study aimed to assess the effects of intravenous \u003csup\u003e64\u003c/sup\u003eZn-asp (KLS-1) administration on systemic inflammation and gut microbiota in a rat model of PD induced by LPS.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eTest Agent\u003c/h2\u003e\u003cp\u003eThe therapeutic agent used in this study was isotopically modified \u003csup\u003e64\u003c/sup\u003eZn di-aspartate (\u003csup\u003e64\u003c/sup\u003eZn-asp), an investigational zinc aspartate complex. The molecule consisted of one atom of \u003csup\u003e64\u003c/sup\u003eZn chelated with two L-aspartic acid molecules (NeoFroxx, Einhausen, Germany), with the molecular formula C\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e64\u003c/sup\u003eZn and a molar mass of 328 g/mol. Zinc accounted for 17.98% of the compound by weight, with the zinc component enriched to 99.2% \u003csup\u003e64\u003c/sup\u003eZn. The compound, designated KLS-1, was synthesized by Pharmaceutical Factory Biopharma LLC (Bila Tserkva, Ukraine).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eAnimals, Experimental Design, and LPS-Induced Parkinson’s Disease Model\u003c/h3\u003e\n\u003cp\u003eThe study was conducted using adult male Wistar rats (8 weeks old, weighing 220\u0026ndash;250 g) obtained from the vivarium of the Educational and Scientific Centre \u0026ldquo;Institute of Biology and Medicine\u0026rdquo; at Taras Shevchenko National University of Kyiv, Ukraine. Animals were housed under standard laboratory conditions with unrestricted access to food and water (standard chow). All animal experiments in this study were performed in accordance with the ARRIVE guidelines.\u003c/p\u003e\u003cp\u003eA total of 51 rats were randomly assigned into four experimental groups using the \u0026ldquo;RAND()\u0026rdquo; function in Microsoft Excel (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e):\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eGroup I \u0026ndash; intact controls (n\u0026thinsp;=\u0026thinsp;12), did not underwent any manipulations;\u003c/p\u003e\u003cp\u003eGroup II \u0026ndash; sham-operated controls (n\u0026thinsp;=\u0026thinsp;12), intra-nigral H\u003csub\u003e2\u003c/sub\u003eOdd injection;\u003c/p\u003e\u003cp\u003eGroup III \u0026ndash; LPS-induced PD model (n\u0026thinsp;=\u0026thinsp;12), intra-nigral LPS injection (10 \u0026micro;g);\u003c/p\u003e\u003cp\u003eGroup IV \u0026ndash; LPS-induced PD model treated with \u003csup\u003e64\u003c/sup\u003eZn-asp (n\u0026thinsp;=\u0026thinsp;15), intra-nigral LPS injection (10 \u0026micro;g)\u0026thinsp;+\u0026thinsp;10 daily i.v. \u003csup\u003e64\u003c/sup\u003eZn-asp injections (1.5 mg/kg).\u003c/p\u003e\u003cp\u003eTo induce unilateral lesions of the nigrostriatal pathway in groups III and IV, stereotaxic injections of 10 \u0026micro;g lipopolysaccharide (LPS, \u003cem\u003eEscherichia coli\u003c/em\u003e O111:B4, Sigma) dissolved in 2 \u0026micro;L sterile saline (JSC \u0026ldquo;Infusion\u0026rdquo;, Ukraine) were administered directly into the substantia nigra as described previously [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Animals in group II received equivalent injections of sterile saline only. Prior to surgery, rats in groups II\u0026ndash;IV were anesthetized with a combination of ketamine (75 mg/kg, Sigma, USA) and 2% xylazine (400 \u0026micro;L/kg, Alfasan International BV, Netherlands), and positioned in a stereotaxic apparatus (SEJ-4, Ukraine). Coordinates for injection were based on Hoban et al. (2013): AP \u0026minus;\u0026thinsp;5.3 mm, ML\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0 mm from bregma, and DV \u0026minus;\u0026thinsp;7.2 mm below the dura [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. The injection rate was 1 \u0026micro;L/min, with the needle left in place for 5 minutes post-injection to facilitate diffusion and prevent backflow.\u003c/p\u003e\u003cp\u003eAn 8-day period post-injection was allowed for disease development. Starting on Day 9, rats in group IV received daily intravenous injections of \u003csup\u003e64\u003c/sup\u003eZn-asp (1.5 mg/kg) via the lateral tail vein for 10 consecutive days. Parkinsonian pathology was confirmed through behavioral assessments and post-mortem analysis of the nigrostriatal system using semi-quantitative tyrosine hydroxylase (TH) immunohistochemistry (data are not presented). On day 28, the rats were sacrificed using carbon dioxide inhalation and subsequent cervical dislocation [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], after which biological samples\u0026mdash;including brain tissue, thymus, peritoneal lavage fluid, sections of the gastrointestinal tract, and blood\u0026mdash;were obtained for analysis.\u003c/p\u003e\n\u003ch3\u003eEthics statement\u003c/h3\u003e\n\u003cp\u003eAll experimental procedures were approved by Ethics Committee of the Taras Shevchenko National University of Kyiv (protocol No. 4, 10.10.2021) and complied with the Animal Welfare Act, as well as national (Kyiv National Bioethics Congress, 2001\u0026ndash;2007) and international (EU Directive 86/609/EEC) guidelines on the ethical treatment of laboratory animals.\u003c/p\u003e\n\u003ch3\u003eBehavioral Assessments\u003c/h3\u003e\n\u003cp\u003eBehavioral testing was conducted to evaluate locomotion, anxiety-like behavior, and dopaminergic dysfunction associated with the LPS-induced Parkinson\u0026rsquo;s disease model.\u003c/p\u003e\n\u003ch3\u003eOpen Field Test\u003c/h3\u003e\n\u003cp\u003eOn Day 24 post-surgery, spontaneous locomotor activity and anxiety-related behavior were assessed using the Open Field test, following the methodology adapted from [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. The test arena consisted of a square enclosure (100 \u0026times; 100 cm) with 30 cm high walls, illuminated by two overhead 60W LED lamps positioned 2 meters above the floor. A 6 \u0026times; 6 grid (36 squares) was marked on the floor to aid spatial tracking. Each rat was individually placed in the center of the arena and allowed to explore for 5 minutes. Movements were recorded from above using a digital camera (Casio\u0026reg; EX-Z850, China) mounted 1 meter above the arena. Video data were later analyzed using MATLAB software. Measured parameters included total distance traveled, time spent in the central zone (inner perimeter), thigmotaxis (time spent near walls), number of rearings and grooming episodes, and frequency of defecation [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. The arena was cleaned and dried between trials to eliminate olfactory cues.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eApomorphine-Induced Rotation Test\u003c/h2\u003e\u003cp\u003eThis test was used to evaluate the extent of dopaminergic neuron loss. It was conducted on Days 8 and 21 following stereotaxic surgery. Apomorphine hydrochloride (Sigma, USA) was administered intraperitoneally at a dose of 0.5 mg/kg. Five minutes after injection, each rat was placed in a cylindrical observation chamber (40 cm diameter), and contralateral (counterclockwise) rotations were recorded manually for 30 minutes using a stopwatch. Rats displaying more than 6 full-body turns per minute (rpm) were considered to have severe dopaminergic neuron (DN) loss (approximately 86.6%), while animals with \u0026le;\u0026thinsp;2 rpm were classified as having moderate DN loss (~\u0026thinsp;44%) [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eElevated Plus Maze (EPM) Test\u003c/h3\u003e\n\u003cp\u003eAnxiety-like behavior was further assessed using the Elevated Plus Maze on Day 24 post-surgery, in accordance with Walf and Frye (2007) [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. The maze consisted of a cross-shaped platform elevated 50 cm above the floor, featuring two open arms (50 \u0026times; 10 cm) and two enclosed arms (50 \u0026times; 10 \u0026times; 30 cm), all connected by a central square (10 \u0026times; 10 cm). Rats were placed in the center square facing an open arm at the start of the 5-minute test session. The setup was illuminated using two ceiling-mounted 60W LED lamps. Behavioral activity was recorded via IP camera and analyzed using MATLAB. Key parameters included total distance traveled, frequency of transitions between arms (open to closed and vice versa), time spent in open vs. closed arms, number of risk assessment behaviors (stretched attend postures), and total number of arm entries. All animals were given a short habituation period to minimize stress before testing.\u003c/p\u003e\n\u003ch3\u003eCRP Level Measurement\u003c/h3\u003e\n\u003cp\u003eC-reactive protein (CRP) levels in blood plasma, collected using the anticoagulant EDTA, were measured by enzyme-linked immunosorbent assay (ELISA) with the ELISA-CRP test system (Labcare Diagnostics India Pvt Ltd), following the manufacturer\u0026rsquo;s instructions.\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eLymphoid organ assessment\u003c/h2\u003e\u003cp\u003eThymus and spleen tissues were carefully excised, weighed, and prepared for a single-cell suspension. Each thymus (spleen) was placed onto a 200-mesh sieve and gently homogenized using a tissue grinder in cold phosphate-buffered saline (PBS, pH 7.3) containing 2% heat-inactivated fetal calf serum (FCS; Gibco, Grand Island, NY, USA) until no visible clumps remained. The resulting single-cell suspensions were then counted using a standard hemocytometer. Cell viability, assessed via Trypan blue exclusion, consistently exceeded 95%. The measured parameters included the relative weights of the thymus and spleen, as well as the relative numbers of thymocytes and splenocytes. These values were calculated as follows:\u003c/p\u003e\u003cp\u003eRelative organ weight = (absolute organ weight/animal weight) x 10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\u003cp\u003eRelative cell count = (absolute cell count / absolute organ weight) x 10\u003csup\u003e\u0026minus;\u0026thinsp;10\u003c/sup\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eHematological Analysis\u003c/h2\u003e\u003cp\u003eWhole blood samples anticoagulated with EDTA were analyzed to assess hematological parameters. A fully automated hematology analyzer (Particle Counter Model PCE 210, ERMA, Japan), calibrated for rodent blood profiling, was used to evaluate blood cell indices in rats. Derived immune-inflammatory ratios were calculated using standard formulas based on absolute cell counts:\u003c/p\u003e\u003cp\u003eNeutrophil-to-lymphocyte ratio (NLR)\u0026thinsp;=\u0026thinsp;absolute neutrophil count, ANC / absolute lymphocyte count, ALC\u003c/p\u003e\u003cp\u003eLymphocyte-to-monocyte ratio (LMR)\u0026thinsp;=\u0026thinsp;ALC / absolute monocyte count, AMC\u003c/p\u003e\u003cp\u003ePlatelet-to-lymphocyte ratio (PLR)\u0026thinsp;=\u0026thinsp;absolute platelet count, APC / ALC\u003c/p\u003e\u003cp\u003ePlatelet-to-neutrophil ratio (PNR)\u0026thinsp;=\u0026thinsp;APC / ANC\u003c/p\u003e\u003cp\u003ePlatelet-to-monocyte ratio (PMR)\u0026thinsp;=\u0026thinsp;APC / AMC\u003c/p\u003e\u003cp\u003eNeutrophil-to-monocyte ratio (NMR)\u0026thinsp;=\u0026thinsp;ANC / AMC\u003c/p\u003e\u003cp\u003eSystemic Immune-Inflammation Index (SII) = (ANC \u0026times; APC) / ALC\u003c/p\u003e\u003cp\u003eSystemic Inflammatory Response index (SIRI) = (ANC x AMC)/ ALC\u003c/p\u003e\u003cp\u003eNeutrophil-platelet-to-lymphocyte\u0026ndash;hemoglobin ratio (NPLHb) = (ANC \u0026times; APC) / (ALC \u0026times; Hb, g/dL)\u003c/p\u003e\u003cp\u003eThese indices were used to evaluate systemic inflammatory status and immune response patterns in the experimental groups.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003ePeritoneal macrophage isolation\u003c/h2\u003e\u003cp\u003ePeritoneal macrophages (PMs) were harvested from the abdominal cavity of non-sensitized mice using a standard rapid peritoneal lavage technique, as previously described [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. The peritoneal cavity was flushed with phosphate-buffered saline (PBS) supplemented with 100 U/mL heparin and 3% fetal bovine serum (FBS) to prevent clotting and preserve cell viability. The collected lavage fluid was centrifuged at 300 \u0026times; g for 5 minutes at 4\u0026deg;C. The resulting cell pellet was washed twice with Hanks\u0026rsquo; Balanced Salt Solution (HBSS) to remove residual serum and debris.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003eAssessment of Phagocyte Metabolic Profile\u003c/h2\u003e\u003cp\u003eThe functional status of phagocytes was evaluated by analyzing their phagocytic capacity, oxidative metabolism, and expression of surface phenotypic markers using flow cytometry. Phagocytosis was assessed based on a previously established method [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. For this purpose, FITC-conjugated, heat-inactivated Staphylococcus aureus Cowan I (sourced from the microbiological collection of the Department of Microbiology and Immunology, ESC \"Institute of Biology and Medicine\", Taras Shevchenko National University of Kyiv) served as the phagocytic target. A suspension containing 2\u0026times;10\u003csup\u003e5\u003c/sup\u003e microglia or macrophages was incubated with 5 \u0026micro;L of the bacterial stock (1\u0026times;10\u003csup\u003e7\u003c/sup\u003e cells/mL) at 37\u0026deg;C for 30 minutes. Phagocytic activity was halted by adding a quenching solution composed of PBS with 0.02% EDTA and 0.04% paraformaldehyde.\u003c/p\u003e\u003cp\u003ePhagocytic efficiency was quantified in two ways: the phagocytosis index (PhI), representing the average fluorescence intensity per phagocytic cell (reflecting the number of bacteria ingested), and the phagocytosis percentage (PP), denoting the proportion of fluorescently positive cells.\u003c/p\u003e\u003cp\u003eReactive oxygen species (ROS) generation, indicative of oxidative metabolic activity, was assessed using the fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA, Invitrogen) as previously described [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTo determine the phenotypic characteristics of phagocytes, cells were stained with FITC-conjugated anti-CD86, PE-conjugated anti-CD80, and Alexa Fluor 647-conjugated anti-CD206 antibodies (BD Pharmingen, USA). All samples were analyzed using a FACSCalibur flow cytometer, and data were processed with CellQuest software (BD Biosciences, USA).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003eFecal water content assessment\u003c/h2\u003e\u003cp\u003eFecal water content was determined using a modified method of Zhu et al. (2012) [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Each rat was placed individually in a clean cage lined with filter paper. Freshly expelled fecal pellets were collected immediately and placed in sealed tubes. The total sample was weighed to obtain the wet weight (WW), then dried at 60\u0026deg;C for 24 h and reweighed to determine the dry weight (DW). Stool water content (%) was calculated using the formula:\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003eSW\u0026thinsp;=\u0026thinsp;100 \u0026minus; (DW \u0026times; 100 / WW)\u003c/h2\u003e\u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\u003ch2\u003eAnalysis of Luminal and Mucosa-Associated Gut Microbiota\u003c/h2\u003e\u003cp\u003eMicrobiota analysis was performed using conventional culture-based techniques as described earlier [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. To evaluate both luminal and wall-adherent microbial populations, distinct sections of the gastrointestinal tract were sampled. For mucosal microbiota assessment, colon segments measuring 1 cm\u003csup\u003e2\u003c/sup\u003e (located 2 cm from the anal verge) and sections of the small intestine (located 2 cm from the ileocecal valve) were excised, washed three times with chyme in saline, and homogenized using a Potter homogenizer. For luminal microbiota analysis, fecal samples were weighed and homogenized in 9 mL of sterile 0.5% sodium chloride solution to obtain a 10\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e dilution. Serial tenfold dilutions (ranging from 10\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e to 10\u003csup\u003e\u0026minus;\u0026thinsp;11\u003c/sup\u003e) were prepared using the same procedure.\u003c/p\u003e\u003cp\u003eAliquots (10 \u0026micro;L) from each dilution were aseptically inoculated onto selective and differential culture media (HiMedia Laboratories Pvt. Ltd., India), including Bifidobacterium Agar, MRS Agar, Endo Agar, Mannitol Salt Agar, Iron Sulphite Agar, Simmons Citrate Agar, and Blood Agar Base (supplemented with 5% sterile defibrinated sheep blood). Cultures were incubated at 37\u0026deg;C for 24\u0026ndash;48 hours.\u003c/p\u003e\u003cp\u003eMicrobial identification was conducted following the taxonomic keys provided in Bergey\u0026rsquo;s Manual of Determinative Bacteriology. Colony morphology, Gram staining, and a battery of biochemical tests were used for classification, including plasma coagulation, DNAse activity, lysozyme and phosphatase production, oxidase activity, carbohydrate fermentation profiles, Voges-Proskauer reaction, motility assessment, and novobiocin susceptibility (to differentiate \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eS. epidermidis\u003c/em\u003e, and \u003cem\u003eS. saprophyticus\u003c/em\u003e). Lactose-negative \u003cem\u003eE. coli\u003c/em\u003e strains were distinguished from other opportunistic \u003cem\u003eEnterobacteriaceae\u003c/em\u003e by their ability to produce hydrogen sulfide.\u003c/p\u003e\u003cp\u003eResults are expressed as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD in logarithmic colony-forming units per gram of feces (lg CFU/g) and per square centimeter of intestinal tissue (lg CFU/cm\u003csup\u003e2\u003c/sup\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eStatistical analysis was performed using the Statistica 12.0 software. The Shapiro\u0026ndash;Wilk test was applied to assess the normality of data distribution, following the approach described by Mishra et al. (2019) [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. Variables with a normal distribution were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD), while those not normally distributed were reported as medians with interquartile ranges (IQR). For group comparisons, normally distributed data were analyzed using one-way ANOVA followed by Tukey\u0026rsquo;s post-hoc test for multiple comparisons. Non-normally distributed data were evaluated using the Mann\u0026ndash;Whitney U test for pairwise comparisons and the Kruskal\u0026ndash;Wallis test for comparisons across multiple groups, as outlined by Chan and Walmsley (1997) [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. The Fisher\u0026rsquo;s Exact Test was employed to compare the proportions of rats showing an increase or decrease in rotation rate between the first and second apomorphine-induced rotation tests. A p-value of \u0026le;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cem\u003eAdministration of\u003c/em\u003e \u003csup\u003e\u003cem\u003e64\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eZn-aspartate attenuated dopaminergic system damage and improved behavioral outcomes in rats with LPS-induced Parkinson\u0026rsquo;s disease\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe apomorphine-induced rotation test is widely recognized as a standard method for evaluating dopaminergic system impairment and behavioral deficits in rat models of PD, including LPS-induced models [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. On Day 8 post-surgery (prior to the initiation of treatment), rats administered LPS exhibited an average contralateral turning rate of 2.2 rpm, which corresponds to an estimated 44\u0026ndash;60% loss of dopaminergic neurons (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). By Day 15, 67% of LPS-lesioned rats demonstrated a 23% increase in rotation rate, suggesting continued neurodegeneration. The remaining 33% of animals in this group showed either stable or slightly reduced turning behavior. In contrast, among the rats receiving \u003csup\u003e64\u003c/sup\u003eZn-asp, 86% showed a 13% decrease in rotation rate by Day 15, indicating potential neuroprotection or partial recovery of dopaminergic function. Only 14% of the treated animals displayed stable or slightly increased rotation rates, suggestive of ongoing neuronal loss.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBehavioral characteristics of rats with LPS-induced Parkinson\u0026rsquo;s disease treated with \u003csup\u003e64\u003c/sup\u003eZn-asp\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIntact animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSham-operated animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLPS-induced PD, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLPS-induced PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp, n\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eOpen field test\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTotal distance traveled, sm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3088.6\u003c/p\u003e\u003cp\u003e[2823.7; 3473.3]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2959.7\u003c/p\u003e\u003cp\u003e[2702.4; 4759.5]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2097.3\u003c/p\u003e\u003cp\u003e[1232.4; 2498.9]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2679.6\u003c/p\u003e\u003cp\u003e[1521.3; 3518.7]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTime spent exploring the inner perimeter, sec\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e21.5\u003c/p\u003e\u003cp\u003e[9.3; 29.5]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e14.0\u003c/p\u003e\u003cp\u003e[7.0; 37.0]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e5.0\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[0.5; 12.0]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.5\u003c/p\u003e\u003cp\u003e[7.3; 22.3]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTime spent in squares surrounded by two walls, min\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.91\u003c/p\u003e\u003cp\u003e[0.90; 0.97]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.95\u003c/p\u003e\u003cp\u003e[0.87; 0.96]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e1.0\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[0.98; 1.1]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea b\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.98\u003c/p\u003e\u003cp\u003e[0.95; 1.0]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber of rearings\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18.8 [16.5; 25.1]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e21.3 [16.0; 28.3]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15.8 [10.3; 21.9]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e16.2 [11.9; 23.3]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRearing duration, s\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.0 [12.5; 18.5]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.8 [8.9; 18.8]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.0 [13.9; 14.3]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.8 [11.9; 16.8]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber of grooming episodes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.0 [2.8; 6.3]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.4 [4.4; 9.3]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e8.8 [6.5; 9.3]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.2 [4.0; 7.3]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber of defecations\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.7 [1.8; 6.4]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.5 [1.9; 6.7]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.8 [1.8; 7.2]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.2 [2.3; 8.4]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eElevated Plus Maze (EPM) Test\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTotal distance traveled, sm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1214.1\u003c/p\u003e\u003cp\u003e[1112.7; 1282.7]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1216.7\u003c/p\u003e\u003cp\u003e[957.6; 1256.0]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e481.04\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[401.3; 635.0]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea b\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e999.7\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[728.3; 1200.3]\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber of transitions\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14.8 [10.8; 18.9]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e16.6 [9.4; 21.6]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10.1 [6.2; 12.4]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.6 [10.2; 19.9]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTime in closed arms/time in open arms\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.1 [2.8; 6.1]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e9.9 [5.8; 10.1]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e15.2\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[10.8; 18.3]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea b\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e11.9 [7.8; 13.1]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTime spent in a stretched attend posture (risk assessment), s\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e200.3\u003c/p\u003e\u003cp\u003e[187.3; 326.1]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e283.6\u003c/p\u003e\u003cp\u003e[211.2; 324.5]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e61.6\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[57.2; 115.1]\u003c/b\u003e \u003csup\u003e\u003cb\u003ea b\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e185.6\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e[135.9; 234.9]\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber of rearings\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18.0 [17.0; 28.0]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25.0 [17.5; 30.5]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e13.5 [9.3; 17.3]\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e16.7 [10.3; 22.5]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eApomorphine test\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eContralateral rotation rate, rpm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDay 8 post lesion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.23 [0.98; 1.65]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.53 [1.23; 1.79]\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDay 21 post lesion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.90 [1.28; 2.38]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e0.85 [0.46; 1.23]\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePercentage of rats exhibiting an increase/ decrease in rotation rate between the first and second apomorphine-induced rotation tests\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e67/33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e14/86\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"5\"\u003eNote: data are presented as median and IQR or %. Data from different animal groups were compared using Kruskal-Wallis\u0026rsquo;s test or Fisher Exact Test for % correspondingly. \u003csup\u003ea\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to intact animals; \u003csup\u003eb\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to sham-operated animals; \u003csup\u003ec\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to untreated animals with LPS-induced PD. Gray shading in the cells indicates statistically significant differences.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eMotor function was assessed using the open field and EPM tests. LPS-induced dopaminergic damage was associated with impaired locomotion (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In the open field test, LPS-lesioned rats exhibited a 37% reduction in the median distance traveled compared to controls (p\u0026thinsp;=\u0026thinsp;0.06). Similarly, in the EPM test, the distance traveled was reduced by approximately 2.4-fold relative to control animals (p\u0026thinsp;\u0026le;\u0026thinsp;0.05). Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp ameliorated these deficits: rats in the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group demonstrated higher locomotor activity in both tests compared to untreated LPS-lesioned animals, with performance levels comparable to those of control groups.\u003c/p\u003e\u003cp\u003eBy the study\u0026rsquo;s end, LPS-lesioned rats exhibited moderate anxiety-like behavior. In the open field test, these animals spent about 2.5 times less time in the central area compared to controls (p\u0026thinsp;\u0026le;\u0026thinsp;0.05), indicating increased anxiety (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The number of transitions between open and closed arms in the EPM was moderately reduced in LPS-lesioned rats. Conversely, LPS-lesioned rats treated with \u003csup\u003e64\u003c/sup\u003eZn-asp tended to show increased transitions, suggesting reduced anxiety. Additionally, the ratio of time spent in closed arms to time in open arms was three times higher in LPS-lesioned rats than in controls, a pattern that was reversed with \u003csup\u003e64\u003c/sup\u003eZn-asp treatment.\u003c/p\u003e\u003cp\u003eThigmotactic behavior, reflected by the time spent in corner zones (squares bordered by two walls), was slightly elevated in LPS-lesioned rats. Furthermore, these rats displayed reduced risk assessment behavior, as evidenced by a more than twofold reduction in the time spent in a stretched-attend posture compared to controls. Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp normalized these parameters, bringing them in line with those observed in healthy rats.\u003c/p\u003e\u003cp\u003eRearing frequency, another indicator of exploratory behavior, was also reduced in LPS-lesioned animals. A tendency toward recovery of this behavior was observed in the treated group. Excessive grooming \u0026ndash; commonly associated with anxiety [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e] \u0026ndash; was prominent in the LPS-PD group but normalized following \u003csup\u003e64\u003c/sup\u003eZn-asp administration.\u003c/p\u003e\u003cp\u003eCollectively, these findings suggest that \u003csup\u003e64\u003c/sup\u003eZn-aspartate treatment not only protects against dopaminergic neurodegeneration but also alleviates motor impairments and mitigates anxiety-like behavior in the LPS-induced PD rat model.\u003c/p\u003e\u003cp\u003e\u003csup\u003e\u003cem\u003e64\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eZn-Aspartate Treatment Improves Hematological Inflammatory Profiles in LPS-Induced Parkinsonian Rats\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe serum level of C-reactive protein (CRP), a widely recognized and reliable marker of systemic inflammation, exhibited substantial individual variability across all experimental groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Despite this variability, the median CRP level in the LPS-PD group was approximately 35% higher than that in control animals. In contrast, the median CRP level in the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group was comparable to that of the control groups.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eWhite blood cell (WBC) counts and their differential components \u0026ndash; neutrophils, lymphocytes, monocytes, and platelets \u0026ndash; are well-established markers of systemic inflammation. In our study, LPS-lesioned rats exhibited a significant increase in both absolute and relative granulocyte (Gr) counts, a decrease in the relative lymphocyte (Ly) count, and a 1.7-fold increase in platelet (PLT) count (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). These changes are indicative of a systemic inflammatory response. In contrast, LPS-lesioned animals treated with \u003csup\u003e64\u003c/sup\u003eZn-aspartate showed no significant differences in these parameters compared to intact and sham-operated control groups, suggesting an anti-inflammatory effect of the treatment.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eHematological parameters in rats with LPS-induced Parkinson\u0026rsquo;s disease treated with \u003csup\u003e64\u003c/sup\u003eZn-asp\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIntact animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSham-operated animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLPS-induced PD, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLPS-induced PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp, n\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e\u003cp\u003eWBC count with differentials\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWBC, x 10^3/\u0026micro;l\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLy, x 10^3/\u0026micro;l\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMo, x 10^3/\u0026micro;l\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGr, x 10^3/\u0026micro;l\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePLT, x 10^3/\u0026micro;l\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e140.3\u0026thinsp;\u0026plusmn;\u0026thinsp;29.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e173.5\u0026thinsp;\u0026plusmn;\u0026thinsp;60.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e240.6\u0026thinsp;\u0026plusmn;\u0026thinsp;24.1 \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e160.6\u0026thinsp;\u0026plusmn;\u0026thinsp;55.8 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLy, %\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e73.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e68.0\u0026thinsp;\u0026plusmn;\u0026thinsp;4.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e60.5\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5 \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e70.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMo, %\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGr, %\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18.2\u0026thinsp;\u0026plusmn;\u0026thinsp;4.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20.0\u0026thinsp;\u0026plusmn;\u0026thinsp;3.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e36.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.4 \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e23.5\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e\u003cp\u003eWBC-based indices of systemic inflammation\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNLR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.25 [0.19; 0.29]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.26 [0.25; 0.30] \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.54 [0.41; 0.62] \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.34 [0.27; 0.39] \u003csup\u003ea b c\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLMR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e9.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePLR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e37.7\u0026thinsp;\u0026plusmn;\u0026thinsp;8.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e45.8\u0026thinsp;\u0026plusmn;\u0026thinsp;11.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e51.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e37.9\u0026thinsp;\u0026plusmn;\u0026thinsp;11.3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePMR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e347.8\u0026thinsp;\u0026plusmn;\u0026thinsp;16.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e253.7\u0026thinsp;\u0026plusmn;\u0026thinsp;34.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e614.5\u0026thinsp;\u0026plusmn;\u0026thinsp;98.7\u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e534.3\u0026thinsp;\u0026plusmn;\u0026thinsp;94.7\u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePNR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e159.8\u0026thinsp;\u0026plusmn;\u0026thinsp;47.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e158.6\u0026thinsp;\u0026plusmn;\u0026thinsp;45.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e152.5\u0026thinsp;\u0026plusmn;\u0026thinsp;67.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e116.4\u0026thinsp;\u0026plusmn;\u0026thinsp;41.6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNMR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eb c\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e38.0 [18.3; 51.6]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e39.1 [33.4; 48.6]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e76.6 [75.9; 78.7] \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e49.0 [41.9; 57.9] \u003csup\u003ea b c\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSIRI\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.11 [0.06; 0.15]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.15 [0.13; 0.19]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.22 [0.19; 0.23] \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.10 [0.07; 0.12] \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNPLHbR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.58 [1.97; 3.15]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.46 [3.03; 4.10]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.73 [5.29; 8.80] \u003csup\u003ea b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.24 [2.87; 3.53] \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"5\"\u003eNote: data are presented as median and IQR or as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Data from different animal groups were compared using Kruskal-Wallis\u0026rsquo;s test or ANOVA with Tukey post-hoc test, respectively. \u003csup\u003ea\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to intact animals; \u003csup\u003eb\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to sham-operated animals; \u003csup\u003ec\u003c/sup\u003e - p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 as compared to untreated animals with LPS-induced PD. Ly \u0026ndash; lymphocytes, Mo \u0026ndash; monocytes, Gr \u0026ndash; granulocytes, PLT \u0026ndash; platelets.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWBC-based indices are now recognized as valuable markers for assessing the severity of systemic inflammation in various conditions, including neurodegenerative diseases. Our study demonstrated a marked increase in nearly all calculated WBC-based inflammatory indices in LPS-PD rats (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In LPS-lesioned rats, NLR was increased 2.2-fold as compared to controls. The median PLR value was elevated by 27.2%. Compared to other inflammatory conditions, PMR and NMR are less commonly used in neurodegenerative disease research, yet they are considered informative markers of systemic inflammation. In our study, both PMR and NMR median values were approximately 2-fold elevated in LPS-lesioned rats. The median SII value was approximately 2-fold higher in the LPS-PD group compared to control animals, while another complex WBC-based index \u0026ndash; SIRI \u0026ndash; was 1.7 times higher than in controls. The median value of a newly proposed WBC-based inflammatory marker, NPLHbR, in the LPS-PD group was approximately 1.9 times higher than that in the control rats. Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp effectively prevented LPS-induced alterations in WBC-based inflammatory indices, maintaining values at levels comparable to those of intact and sham-operated controls, unlike in untreated LPS-PD rats.\u003c/p\u003e\u003cp\u003e\u003csup\u003e\u003cem\u003e64\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eZn-asp Modulates Polarization of Peripheral Blood Phagocytes in LPS-Induced PD Rats\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe functional state of circulating phagocytes was assessed using parameters commonly applied to characterize their polarized activation profile: phagocytic activity (PI), oxidative metabolism (ROS generation), and the expression of phenotypic markers CD80/86 and CD206. In the LPS-PD group, the median monocyte PI value was twice lower than that of control animals (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). Treatment of LPS-lesioned rats with \u003csup\u003e64\u003c/sup\u003eZn-asp increased the monocyte PI median by 31% compared to untreated parkinsonian rats, with values comparable to those in the sham-operated control group. The median neutrophil PI value in the LPS-PD group did not differ significantly from that of control rats, and treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp had no effect on this parameter (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Notably, neutrophil phagocytic activity was elevated in sham-operated animals, likely reflecting an N2 polarization shift associated with their participation in reparative processes following surgical intervention [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]. Monocyte ROS generation was markedly elevated in LPS-lesioned rats, indicating a pro-inflammatory shift characteristic of systemic inflammation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp substantially reduced oxidative metabolism: the median ROS generation in the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group was sevenfold lower than in the LPS-PD group and approximately fourfold lower than in controls. Neutrophil ROS generation in LPS-PD rats was significantly higher than in controls. In contrast, LPS-lesioned animals treated with \u003csup\u003e64\u003c/sup\u003eZn-asp showed ROS levels comparable to sham-operated rats (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). Notably, sham-operated rats exhibited reduced ROS generation compared to intact animals, further supporting the notion of an anti-inflammatory, tissue-repairing metabolic profile. Neither the proportion of CD80/86⁺ cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE) nor the CD80/86 expression level (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF) in circulating phagocytes from LPS-lesioned rats differed significantly from controls. In the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group, the median proportion of CD80/86⁺ cells was reduced by half compared with untreated animals, while the expression level was approximately threefold higher. The proportion of CD206⁺ circulating phagocytes was slightly elevated in LPS-lesioned rats, whereas treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp restored this value to control levels (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eG). CD206 expression level in the LPS-PD group was similar to intact animals but 6.5-fold lower than in sham-operated rats (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eH). Administration of \u003csup\u003e64\u003c/sup\u003eZn-asp modestly increased CD206 expression. Since elevated CD206 is linked to an anti-inflammatory phagocyte phenotype, the higher values in sham-operated rats likely reflect post-surgical reparative processes, while the increase in the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group may result from the drug\u0026rsquo;s anti-inflammatory action.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\u003ch2\u003eInfluence of \u003csup\u003e64\u003c/sup\u003eZn-asp on Lymphoid Organ Weight and Cellularity\u003c/h2\u003e\u003cp\u003ePD-related inflammation was reflected in alterations of the lymphoid organs. LPS-PD rats showed significantly reduced thymus weight and increased thymus cellularity compared with controls (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eLymphoid organ parameters in rats with LPS-induced Parkinson\u0026rsquo;s disease treated with \u003csup\u003e64\u003c/sup\u003eZn-asp\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIntact animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSham-operated animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLPS-induced PD, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLPS-induced PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp, n\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRelative weight of thymus\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.32 [1.27; 1.36]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.26 [1.18; 1.33]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.07 [0.08; 1.08] \u003csup\u003ea,b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.16 [1.07; 1.29] \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRelative number of thymocytes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1.75 [1.49; 2.01]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.77 [1.72; 5.82] \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.86 [6.13; 9.86] \u003csup\u003ea,b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.26 [4.24; 8.31] \u003csup\u003ea,b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRelative weight of spleen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.97 [2.62; 3.33]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.31 [3.07; 3.56]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2.55 [2.48; 2.59] \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.19 [2.32; 3.45] \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRelative number of splenocytes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.68 [3.26; 4.10]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.58 [5.53; 5.63] \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.45 [5.69; 8.62] \u003csup\u003ea,b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.46 [2.44; 4.27] \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eTreatment with \u003csup\u003e64\u003c/sup\u003eZn-asp restored thymus weight to control levels and partially normalized cellularity. Spleen weight was unchanged in LPS-lesioned rats compared to intact controls, but was marginally lower than in sham-operated animals; cellularity showed only a slight increase. In the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group, both spleen weight and splenocyte counts were indistinguishable from controls.\u003c/p\u003e\u003cp\u003e\u003csup\u003e\u003cem\u003e64\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eZn-asp Alters Polarization Profiles of Peritoneal Macrophages in LPS-Induced PD Rats\u003c/em\u003e\u003c/p\u003e\u003cp\u003ePeritoneal macrophage PI was comparable across all experimental groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). In LPS-PD rats, median ROS generation was approximately twice that of intact and sham-operated controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB), indicating heightened oxidative activity. Administration of \u003csup\u003e64\u003c/sup\u003eZn-asp normalized ROS production to control levels. The proportion of CD206⁺ cells (representing large resident peritoneal macrophages) and CD206 expression levels in LPS-lesioned rats were similar to those in intact animals (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC, D). Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp reduced the CD206⁺ cell fraction but increased CD206 expression.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eNeither the percentage of CD80/86⁺ cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE) nor CD80/86 expression levels (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF) in LPS-lesioned rats differed significantly from controls, and \u003csup\u003e64\u003c/sup\u003eZn-asp treatment had no effect on these parameters.\u003c/p\u003e\u003cp\u003e\u003csup\u003e\u003cem\u003e64\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eZn-Asp Mitigates Inflammation-Linked Gut Dysbiosis in a Rat with LPS-induced PD\u003c/em\u003e\u003c/p\u003e\u003cp\u003eWe analyzed the composition of the culturable gut microbiota in conjunction with measurements of fecal water content. At the end of the experiment, stool water content did not differ significantly among the experimental groups (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Nevertheless, rats in the LPS-PD group produced approximately 1.5-fold more feces (wet weight: 0.553\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 g; dry weight: 0.268\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 g) than controls (sham: 0.387\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 g wet; 0.183\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 g dry; p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), indicating impaired colonic motility, with stool accumulating in the colon rather than being efficiently propelled and expelled. In the LPS-PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp group, fecal output was slightly lower than in untreated LPS-PD rats but remained higher than in controls.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eFecal wet weight, dry weight, and stool water content in rats with LPS-induced Parkinson\u0026rsquo;s disease treated with \u003csup\u003e64\u003c/sup\u003eZn-asp\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIntact animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSham-operated animals, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLPS-induced PD, n\u0026thinsp;=\u0026thinsp;12\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLPS-induced PD\u0026thinsp;+\u0026thinsp;\u003csup\u003e64\u003c/sup\u003eZn-asp, n\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFecal wet weight, g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e0.356\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.387\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.554\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 \u003csup\u003ea, b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.521\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 \u003csup\u003ea, b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFecal dry weight, g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e0.191\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.183\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.268\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u003csup\u003ea, b\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.233\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStool water content, %\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e54.1\u0026thinsp;\u0026plusmn;\u0026thinsp;3.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e48.5\u0026thinsp;\u0026plusmn;\u0026thinsp;7.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48.9\u0026thinsp;\u0026plusmn;\u0026thinsp;8.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e47.9\u0026thinsp;\u0026plusmn;\u0026thinsp;6.57\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"5\"\u003eNotes: Data are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Data from different animal groups were compared using ANOVA with a Tukey post-hoc test. a - p\u0026thinsp;\u0026le;\u0026thinsp;0.05 as compared to intact animals; b - p\u0026thinsp;\u0026le;\u0026thinsp;0.05 as compared to sham-operated animals.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eIn gut microbiota assessment, particular attention was focused on quantitative analysis of anaerobic saccharolytic genera \u003cem\u003eBifidobacterium\u003c/em\u003e and \u003cem\u003eLactobacillus\u003c/em\u003e, which synthesize γ-aminobutyric acid (GABA) involved in gastrointestinal motility and the gut-brain axis. In LPS-PD rats, counts of these bacteria were moderately reduced in the small-intestinal and large-intestinal wall-adherent microbiota (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA) and markedly decreased in the luminal microbiota of the large intestine (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB). Specifically, the number of \u003cem\u003eBifidobacterium\u003c/em\u003e species decreased by one order of magnitude \u0026mdash; from lg 8.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6 CFU/g in sham-operated animals to lg 6.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 CFU/g in the LPS-PD group. The number of \u003cem\u003eLactobacillus\u003c/em\u003e species decreased by two orders of magnitude compared to the control group \u0026mdash; from lg 7.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 CFU/g in controls to lg 5.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.70 CFU/g in the LPS-lesioned rats. Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp preserved bacterial abundance at control levels.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eUnder normophysiological conditions, neither lactose-positive nor lactose-negative \u003cem\u003eE. coli\u003c/em\u003e were detected in the mucosa-associated biotope (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eC). In the LPS-PD group, lactose-positive \u003cem\u003eE. coli\u003c/em\u003e reached 10\u003csup\u003e3\u003c/sup\u003e CFU/cm\u003csup\u003e2\u003c/sup\u003e, while lactose-negative strains increased to 10\u003csup\u003e5\u003c/sup\u003e CFU/cm\u003csup\u003e2\u003c/sup\u003e in the mucosa-associated microbiota of the small intestine, with trace amounts also detected in colonic tissue. LPS-PD was additionally associated with a pronounced increase in opportunistic enterobacteria, reaching lg 2.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 CFU/cm\u003csup\u003e2\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eIn fecal microbiota from LPS-PD animals, total \u003cem\u003eE. coli\u003c/em\u003e abundance did not differ significantly from sham-operated controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD). However, opportunistic enterobacteria counts increased by approximately two orders of magnitude, from lg 2.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73 CFU/g in controls to lg 4.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52 CFU/g in the LPS-PD group.\u003c/p\u003e\u003cp\u003eTreatment with \u003csup\u003e64\u003c/sup\u003eZn-asp resulted in a notable restoration of microbiota composition. Lactose-fermenting \u003cem\u003eE. coli\u003c/em\u003e levels decreased, and lactose-negative strains were completely eliminated from the mucosa-associated microbiota. Moreover, \u003csup\u003e64\u003c/sup\u003eZn-asp administration significantly reduced opportunistic enterobacteria abundance in both mucosa-associated and luminal compartments.\u003c/p\u003e\u003cp\u003e\u003cem\u003eStaphylococcus\u003c/em\u003e spp., including both mannitol-fermenting (\u003cem\u003eS. aureus\u003c/em\u003e) and mannitol-negative strains, were also detected [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. No \u003cem\u003eStaphylococci\u003c/em\u003e were present in the mucosa-associated biotopes of control animals. In LPS-PD animals, \u003cem\u003eStaphylococcus\u003c/em\u003e spp. were identified in the wall-adherent microbiota of the large intestine, suggesting increased aerobiosis (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE). Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp slightly decreased the abundance of mannitol-fermenting staphylococci in the mucosa-associated biotope. Quantitative characteristics of staphylococci in the luminal microbiota did not differ significantly between groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF).\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eLPS-induced PD models are among the most commonly used inflammatory models of PD, as they effectively reproduce key pathological features, including motor impairment, neuroinflammation, and systemic inflammation [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Furthermore, these models provide valuable tools for investigating anti-inflammatory and neuroprotective agents [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Given the growing recognition of systemic inflammation as a contributor to PD development and progression, as well as a potential therapeutic target, we employed this model to evaluate the effects of intravenous \u003csup\u003e64\u003c/sup\u003eZn-asp administration on systemic immune-inflammatory responses in PD rats. Findings from \u003cem\u003ein vitro\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e studies suggest that zinc may exert disease-modifying effects in PD through several mechanisms. Zinc appears to influence autophagy and lysosomal function, which may help limit α-synuclein accumulation, and it can reduce aggregation by enhancing albumin chaperone activity [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. It has also been reported to modulate inflammatory pathways, including NF-κB signaling [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e], the NLRP3 inflammasome [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e], and STAT3 activation [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e], while preserving immune competence [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. In addition, zinc contributes to redox homeostasis by inducing metallothioneins and glutathione, thereby supporting antioxidant defenses [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn our study, intravenous administration of \u003csup\u003e64\u003c/sup\u003eZn-asp exerted neuroprotective effects by reducing dopaminergic neurodegeneration, ameliorating motor dysfunction, and attenuating anxiety-like behavior in Parkinsonian rats. These outcomes were associated with the pronounced anti-inflammatory activity of the drug.\u003c/p\u003e\u003cp\u003eSystemic inflammation in LPS-induced PD rats was confirmed by established markers, including elevated serum CRP levels, increased granulocyte and platelet counts, and a concomitant reduction in lymphocyte count [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. Beyond these parameters, indices derived from complete blood counts provide a more nuanced assessment of immune-inflammatory status by capturing the relative proportions of different leukocyte populations. Among them, the NLR, coupled with relative lymphopenia, has been linked to neurodegeneration-associated protein alterations, particularly within α-synuclein and amyloid-β pathways, and correlates with greater clinical burden in PD patients [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]. The PLR represents another established indicator of peripheral immune dysregulation and systemic inflammation in PD [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e]. Similarly, an elevated NMR suggests an intensified inflammatory response [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. Platelets, in addition to their roles in hemostasis and thrombosis, are increasingly recognized as active mediators of inflammation, supporting the recruitment of lymphocytes, neutrophils, and monocytes to inflamed tissues, thereby amplifying immune responses. In PD, chronic inflammation promotes platelet hyperreactivity, which may further exacerbate neuroinflammation [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. This highlights the relevance of the PMR as an informative marker of systemic inflammatory activity. Composite indices have also been proposed to better capture the complexity of immune-inflammatory dynamics. The SII, which incorporates neutrophil, platelet, and lymphocyte counts, provides an integrated measure of the immune-inflammatory balance. Elevated SII is strongly associated with increased PD risk, particularly in females [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. Likewise, the SIRI, which combines neutrophil, monocyte, and lymphocyte counts, reflects the interplay between immune activation and suppression and may indicate states of immunodeficiency or immune exhaustion [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e]. More recently, the NPLHbR has been introduced as a reliable WBC-based marker of systemic inflammation. Unlike other indices, NPLHbR also incorporates hemoglobin, a parameter frequently reduced in chronic inflammatory states, including age-related \u0026ldquo;inflammaging,\u0026rdquo; which is associated with anemia and sustained immune activation [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e]. In our study, the median values of all WBC-based inflammatory indices were significantly (~\u0026thinsp;2 times) elevated in LPS-lesioned rats, reflecting pronounced and persistent systemic inflammation accompanied by features of immune exhaustion. \u003csup\u003e64\u003c/sup\u003eZn-aspartate prevented LPS-induced changes in WBC-based inflammatory indices, preserving values comparable to controls. In addition to the well-established anti-inflammatory properties of zinc mediated through modulation of key pro-inflammatory signaling pathways such as NF-κB and IL-6, promotion of anti-inflammatory protein expression, and regulation of nitric oxide production [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e], one potential mechanism underlying the observed effects of \u003csup\u003e64\u003c/sup\u003eZn-asp may involve its impact on thymic function. In the context of systemic inflammation, acute thymic atrophy, as observed in LPS-lesioned rats in our study and in the MPTP model of PD in mice [\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e], may occur, leading to extensive loss of developing T cells during thymic selection, along with infiltration of immune cells that can further disrupt thymic architecture [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e]. Zinc plays a pivotal role in normal T-cell development and in recovery following acute injury and inflammation, as it promotes thymic regeneration by inducing endothelial cell production of bone morphogenetic protein 4 (BMP4) [\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e]. In addition, zinc can promote differentiation of regulatory T-cells [\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e], which interfere with inflammatory signaling pathways controlling systemic inflammation [\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn addition to restoring immune cell counts in LPS-PD animals, intravenous administration of \u003csup\u003e64\u003c/sup\u003eZn-asp abrogated the pro-inflammatory metabolic shift of phagocytic cells \u0026ndash; key mediators of inflammatory responses \u0026ndash; in both peripheral blood and the peritoneal cavity. In PD, systemic inflammation is driven in part by monocytes and neutrophils. Clinical studies consistently report elevated circulating levels of these cells along with increased concentrations of inflammatory markers. Among monocytes, the classical pro-inflammatory (M1) subset is particularly implicated, as these cells release cytokines and chemokines that exacerbate neuroinflammation and contribute to neuronal damage. Neutrophils, the most abundant immune cells in circulation, act as early responders to inflammatory stimuli. In PD, pro-inflammatory N1 neutrophils are believed to play a role in the initial immune response to potential triggers, including pathogens or cellular damage. Although their role in PD has been less extensively investigated than that of monocytes and microglia, they are increasingly recognized as important contributors to early inflammatory events [\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e]. In our study, LPS-lesioned rats exhibited an increased proportion of circulating CD206⁺ phagocytic cells, likely representing a subset of activated myeloid cells with a pro-inflammatory metabolic profile. As described by Trombetta et al. (2018), these cells display a mixed phenotype, co-expressing markers of both M1 (pro-inflammatory) and M2 (anti-inflammatory) phagocytes. A further indication of their pro-inflammatory metabolic bias is their reduced maturation relative to counterparts expressing the same M2 markers but displaying a more anti-inflammatory phenotype, as well as the absence of CD204 expression [\u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e82\u003c/span\u003e]. In addition, reduced phagocytic activity in monocytes, together with markedly elevated ROS production in both neutrophils and monocytes of LPS-lesioned rats, further supports the presence of a pro-inflammatory metabolic shift in these cells [\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e83\u003c/span\u003e, \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e84\u003c/span\u003e]. Treatment with \u003csup\u003e64\u003c/sup\u003eZn-asp returned CD206\u0026thinsp;+\u0026thinsp;phagocyte cell fraction to the norm, marginally increased monocyte phagocytic activity, and substantially decreased the oxidative metabolism of both phagocytic cell populations, which indicates an anti-inflammatory metabolic shift. In treated parkinsonian rats, we observed a reduced fraction of CD80/86⁺ cells accompanied by increased CD80/86 expression levels. This paradoxical pattern may indicate a shift toward the differentiation of CD80/86⁺ monocyte-derived myeloid suppressor cells, potentially driven by the drug\u0026rsquo;s anti-inflammatory action. Costimulatory molecules of the B7.1 family, CD80 and CD86, serve a dual role as markers of phagocyte polarization. On the one hand, upregulation of CD80/CD86 expression reflects the acquisition of antigen-presenting capacity by circulating phagocytes and is indicative of a pro-inflammatory metabolic profile [\u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e85\u003c/span\u003e]. On the other hand, the majority of \u003cem\u003eex vivo\u003c/em\u003e-generated myeloid-derived suppressor cells (MDSCs) exhibit high CD80/CD86 expression, and monocytic MDSCs \u003cem\u003ein vivo\u003c/em\u003e are characterized by strong CD86 positivity [\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e, \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe metabolic profile of peritoneal macrophages in LPS-lesioned rats was moderately altered compared with controls, with increased ROS production indicating a pro-inflammatory metabolic shift [\u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e88\u003c/span\u003e]. Treatment with \u003csup\u003e64\u003c/sup\u003eZn-aspartate reduced ROS generation and enhanced CD206 expression, a reliable marker of an anti-inflammatory phenotype in tissue-resident macrophages [\u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e89\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe hypothesis that PD may originate in the periphery and involve environmental risk factors has brought increasing attention to the role of the gut microbiota. Intestinal dysbiosis, which can precede the clinical diagnosis of PD by several years, has been proposed as a potential trigger of the disease through the disruption of microbial and mucosal immune homeostasis, thereby initiating an inflammatory cascade that extends to the brain and contributes to the pathophysiology of PD [\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e]. Nevertheless, the prevailing view is that dysbiosis is more likely a consequence of PD rather than its primary cause. Although it may arise years before clinical onset and contribute to disease progression, it is not generally considered the initiating event [\u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e91\u003c/span\u003e]. Instead, inflammation inherent to PD appears to be a major driver of gut dysbiosis. Under inflammatory conditions, the relative abundance of obligate anaerobes belonging to the \u003cem\u003eBacteroidetes\u003c/em\u003e and \u003cem\u003eFirmicutes\u003c/em\u003e phyla declines, while \u003cem\u003eProteobacteria\u003c/em\u003e \u0026ndash; particularly \u003cem\u003eGammaproteobacteria\u003c/em\u003e such as \u003cem\u003eEnterobacteriaceae\u003c/em\u003e \u0026ndash; expand and become dominant within the gut microbiota [\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e]. In our study, systemic inflammation in parkinsonian rats was closely associated with pronounced intestinal dysbiosis, suggesting a bidirectional interplay between peripheral immune activation and gut microbial imbalance. Representatives of the anaerobic saccharolytic genera \u003cem\u003eBifidobacterium\u003c/em\u003e and \u003cem\u003eLactobacillus\u003c/em\u003e exhibited a marked reduction in both mucosa-associated and luminal microbiota. In parallel, members of the family \u003cem\u003eEnterobacteriaceae\u003c/em\u003e demonstrated a significant expansion. Specifically, both lactose-fermenting and non-fermenting \u003cem\u003eEscherichia coli\u003c/em\u003e, as well as opportunistic enterobacteria, were detected in the wall-adherent microbiota of LPS-lesioned animals, whereas these taxa were absent in control rats. A comparable increase in opportunistic enterobacteria was also observed in the colonic luminal microbiota. Both systemic and intestinal inflammation are known to alter the physiological oxygen gradient, leading to localized increases in oxygen availability. This shift generates conditions that are more aerobic than those observed in the healthy gut. Within this altered niche, members of the \u003cem\u003eEnterobacteriaceae\u003c/em\u003e family gain a selective advantage, while commensal symbiotic bacteria are negatively affected by inflammation-induced environmental changes [\u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e93\u003c/span\u003e]. Among the pathobionts characteristic of the gut microbiota in patients with PD, adherent-invasive \u003cem\u003eE. coli\u003c/em\u003e (AIEC) has been identified. AIEC is thought to contribute to PD through the production of substances such as curli, which promote the aggregation of α-synuclein, a key pathogenic process in the disease [\u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e94\u003c/span\u003e]. Inflammation in the peritoneal cavity, reflected by the pro-inflammatory metabolic profile of peritoneal macrophages and accompanied by pronounced dysbiosis, suggests concurrent inflammation within the intestinal mucosa-associated lymphoid tissue, which may promote the growth and virulence of AIEC. Notably, the emergence of \u003cem\u003eStaphylococcus\u003c/em\u003e, atypical for mucosa-associated intestinal microbiota, likely reflects inflammation-driven aerobization of the biotope, further highlighting microbial community disruption. In LPS-lesioned rats, gut microbiota imbalance was associated with increased fecal output without a corresponding rise in water content, indicating gastrointestinal dysfunction \u0026ndash; one of the most common non-motor symptoms in PD [\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e95\u003c/span\u003e, \u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e96\u003c/span\u003e]. This dysbiosis may also contribute to the thymic atrophy observed in these animals [\u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e97\u003c/span\u003e]. Treatment of LPS-PD rats with \u003csup\u003e64\u003c/sup\u003eZn-asp promoted the rebalancing of gut microbial communities, evidenced by an increase in bifidobacteria and lactobacilli in the luminal microbiota and a concomitant reduction or disappearance of enterobacteria in the wall-adherent compartment, suggesting a microbiota-stabilizing effect. This effect may result from the anti-inflammatory action of the compound, although a direct impact of the preparation on the microorganisms cannot be excluded. Zinc has been reported to exert bidirectional effects on \u003cem\u003eLactobacillus\u003c/em\u003e species, promoting the growth of some while inhibiting others [\u003cspan citationid=\"CR98\" class=\"CitationRef\"\u003e98\u003c/span\u003e]. In addition, zinc can suppress the growth and virulence of AIEC by reducing bacterial adherence, biofilm formation, and the expression of key virulence factors [\u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e99\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThis study is subject to several limitations. First, it did not directly investigate how 64Zn-asp influences zinc homeostasis or neuronal function, leaving important aspects of its underlying mechanism unresolved and emphasizing the need for further exploration. Second, evaluating the metabolic profile of phagocytes requires incorporating additional phenotypic markers to enable more precise differentiation of myeloid-derived suppressor cells. Finally, molecular approaches are necessary to gain deeper insight into the impact of \u003csup\u003e64\u003c/sup\u003eZn-asp on the intestinal microbiota.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eSystemic inflammation is a recognized contributor to the onset and progression of PD, making it an important therapeutic target. Peripheral inflammatory events can intensify neuroinflammation and accelerate neurodegeneration; therefore, anti-inflammatory interventions are being actively explored. In this study, we demonstrated that a zinc preparation with a unique isotopic composition \u003csup\u003e64\u003c/sup\u003eZn-asp \u0026ndash; endowing it with high biological activity and potent anti-inflammatory properties \u0026ndash; was able to attenuate systemic inflammation in animals with LPS-PD while also modulating gut dysbiosis. These findings highlight the potential of this preparation as a multifaceted therapeutic strategy; however, further mechanistic studies and translational research are essential to validate its efficacy and safety in the context of human PD.\u003c/p\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data that support the findings of this study are available from the corresponding author, upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMT: supervision and review. MR: methodology, investigation, formal analysis. AB: methodology, conceptualization, and project administration. SG: conceptualization, writing, review, and editing. TD: methodology, investigation. RB: methodology, conceptualization. ND: methodology, investigation. RD: methodology, investigation. TS: methodology, investigation, and writing original draft. GT: supervision and writing \u0026ndash; review and editing. LS: methodology, conceptualization, project administration, and writing the original draft. All authors read the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was supported by a TSNUK project N 18DP036-10.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eSu, D. et al. \u003cem\u003eProjections for prevalence of Parkinson's disease and its driving factors in 195 countries and territories to 2050: modelling study of Global Burden of Disease Study 2021\u003c/em\u003e Vol. 388, e080952 (BMJ, 2025). 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The micronutrient zinc inhibits EAEC strain 042 adherence, biofilm formation, virulence gene expression, and epithelial cytokine responses, benefiting the infected host. \u003cem\u003eVirulence\u003c/em\u003e \u003cb\u003e4\u003c/b\u003e (7), 624\u0026ndash;633. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4161/viru.26120\u003c/span\u003e\u003cspan address=\"10.4161/viru.26120\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Parkinson’s disease, Inflammation, Gut dysbiosis, Stable light isotope-enriched zinc aspartate, Motor function, Anxiety-like behavior","lastPublishedDoi":"10.21203/rs.3.rs-8178695/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8178695/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA growing body of research indicates that systemic inflammation contributes substantially to the progression of Parkinson\u0026rsquo;s disease (PD). Foundational studies propose that targeting inflammatory pathways may offer therapeutic benefits for PD and other neurodegenerative conditions. Our previous work demonstrated that a novel zinc aspartate compound enriched with the light isotope \u003csup\u003e64\u003c/sup\u003eZn (\u003csup\u003e64\u003c/sup\u003eZn-asp) can counteract inflammatory and cognitive impairments triggered by intra-hippocampal Aβ\u003csub\u003e1\u0026minus;40\u003c/sub\u003e in rats, and can also mitigate neuroinflammation while promoting neuronal survival in a PD model. In the present study, we investigated the impact of this isotopically modified zinc compound on systemic inflammatory responses and gut microbiota composition in a rat model of PD induced by a single stereotactic intranigral injection of lipopolysaccharide (LPS).\u003c/p\u003e\u003cp\u003eLPS-lesioned rats exhibited impaired locomotion, heightened anxiety-like behavior, and progressive dopaminergic dysfunction. \u003csup\u003e64\u003c/sup\u003eZn-asp administration attenuated behavioral deficits and reduced apomorphine-induced rotations. Treatment normalized CRP levels, reversed LPS-induced increases in granulocytes and platelets, and corrected elevations in systemic inflammatory indices (including NLR, PLR, SII, and SIRI). \u003csup\u003e64\u003c/sup\u003eZn-asp shifted circulating and peritoneal phagocytes toward an anti-inflammatory phenotype and partially restored thymus structure and cellularity. In the gut, LPS-induced PD resulted in marked reductions in \u003cem\u003eBifidobacterium\u003c/em\u003e and \u003cem\u003eLactobacillus\u003c/em\u003e spp. and an expansion of opportunistic \u003cem\u003eEnterobacteriaceae\u003c/em\u003e and \u003cem\u003eStaphylococcus\u003c/em\u003e spp. \u003csup\u003e64\u003c/sup\u003eZn-asp largely preserved beneficial anaerobes and suppressed opportunistic taxa in both luminal and mucosa-associated compartments.\u003c/p\u003e\u003cp\u003eThese findings demonstrate that \u003csup\u003e64\u003c/sup\u003eZn-aspartate exerts anti-inflammatory, immunomodulatory, and microbiota-stabilizing effects, suggesting potential therapeutic value as a disease-modifying strategy targeting neuroimmune-gut axis dysfunction in PD.\u003c/p\u003e","manuscriptTitle":"Isotopically Enriched 64Zn-aspartate Attenuates Systemic Inflammation and Gut Dysbiosis in an Lps-induced Rat Model of Parkinson’s Disease","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-15 22:32:29","doi":"10.21203/rs.3.rs-8178695/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-21T06:59:57+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-20T11:02:03+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-15T21:46:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"292713126264850125720933715806728958126","date":"2026-01-09T14:59:04+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"168104306429849338521386821629574272862","date":"2025-12-15T13:28:19+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-10T12:07:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-12-10T11:58:56+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-28T06:49:01+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-26T09:51:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-26T09:38:44+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"33eec25a-a742-4f59-9a9d-ca23aea0542f","owner":[],"postedDate":"December 15th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":59709882,"name":"Health sciences/Diseases"},{"id":59709883,"name":"Biological sciences/Immunology"},{"id":59709884,"name":"Biological sciences/Microbiology"},{"id":59709885,"name":"Health sciences/Neurology"},{"id":59709886,"name":"Biological sciences/Neuroscience"}],"tags":[],"updatedAt":"2026-03-30T16:18:27+00:00","versionOfRecord":{"articleIdentity":"rs-8178695","link":"https://doi.org/10.1038/s41598-026-45640-9","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-03-27 16:11:07","publishedOnDateReadable":"March 27th, 2026"},"versionCreatedAt":"2025-12-15 22:32:29","video":"","vorDoi":"10.1038/s41598-026-45640-9","vorDoiUrl":"https://doi.org/10.1038/s41598-026-45640-9","workflowStages":[]},"version":"v1","identity":"rs-8178695","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8178695","identity":"rs-8178695","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}