Protective Effects of Date Syrup And Green Tea Against Atrazine-Induced Hepatic And Reproductive Toxicity In Male Albino Wistar Rats

Protective Effects of Date Syrup And Green Tea Against Atrazine-Induced Hepatic And Reproductive Toxicity In Male Albino Wistar Rats | 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 Research Article Protective Effects of Date Syrup And Green Tea Against Atrazine-Induced Hepatic And Reproductive Toxicity In Male Albino Wistar Rats Hassan Abdulsalam Adewuyi, Abiola Usman Mohammed, Tolulope Olukayode Jaiyeola, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7416536/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background : Drug-induced hepatotoxicity and reproductive dysfunction remain significant clinical challenges, necessitating novel protective agents. Existing therapies often fail to concurrently address liver and gonadal damage, highlighting the need for multifunctional interventions. Aim : This study evaluated the hepatoprotective and fertility-preserving effects of a novel compound in a atrazine-induced rodent model, comparing its efficacy against standard and experimental treatments Study design: Experimental study. Place and Duration of Study: Department of Pharmacy Technician, Federal College of Health Technology Ilese-Ijebu Ogun State Nigeria, between August 2024 and November 2024.. Methodology : Forty-eight male Wistar rats were divided into six groups (n=5): Group I (normal control), Group II (positive control, toxin-exposed), and Groups III-VI (toxin + test compounds). After 28 days, serum AST, ALT, GGT, ALP, 5'NT, albumin, total protein, sperm parameters, and hormonal levels were analyzed using standard procedures. Statistical significance was assessed via ANOVA and Tukey’s post-hoc tests (p < 0.05). Results : Group II exhibited severe hepatotoxicity (AST: 81.56±1.245 IU/L; ALT: 57.32±0.412 IU/L) and reproductive impairment (sperm count: 22.6±1.8×10⁶/mL; testosterone: 3.45±0.18 ng/mL). Group VI demonstrated near-normal liver enzymes (AST: 70.62±0.495; ALT: 38.94±3.872), preserved albumin (4.660±0.0512 g/dL), and improved sperm count (42.6±2.0×10⁶/mL) versus Group II (p < 0.0001). Conclusion : The novel compound in Group VI significantly mitigated both hepatic and reproductive toxicity, outperforming other interventions. These findings suggest a dual-organ protective mechanism, potentially mediated through antioxidant or anti-inflammatory pathways. Future studies should isolate the active constituents of Group VI and validate results in higher mammalian models. Clinical trials are warranted to assess translational potential in chemotherapy-associated toxicity. Toxicology Clinical Pharmacology Date syrup Green Tea Hepatoprotection reproductive toxicity dual-organ protection therapeutic intervention 1. Introduction Atrazine is a widely used herbicide associated with significant toxicological effects, including oxidative stress, inflammation, and hepatic injury upon chronic exposure [1]. While synthetic pharmaceuticals may offer partial protection against atrazine-induced damage, their clinical utility is often limited by adverse side effects and high costs [2]. Consequently, researchers are increasingly exploring natural alternatives such as date syrup and green tea, which contain bioactive compounds with potential hepatoprotective properties [3]. This study aims to evaluate the efficacy of these natural substances in mitigating atrazine-induced liver damage, providing insights into safer and more sustainable therapeutic approaches. Atrazine is primarily employed in agriculture to control broadleaf weeds in crops such as corn, sugarcane, and sorghum, making it one of the most extensively used herbicides globally [4]. Despite its agricultural benefits, atrazine exhibits environmental persistence, leading to contamination of soil, groundwater, and aquatic ecosystems [5]. Atrazine exposure induces hepatic oxidative stress and reproductive dysfunction in experimental models; polyphenol-rich agents such as green tea and phytochemical fractions from local botanicals have shown protective effects in similar models [6,7,8]. Phytochemical fractionation methodologies inform selection of active fractions (ethyl acetate, aqueous) with antioxidant activity [9,10,11]. The liver is a vital organ responsible for detoxification, metabolic regulation, and maintaining systemic homeostasis [12]. Anatomically, it resides in the right upper abdominal quadrant, comprising two primary lobes (right and left) subdivided into functional lobules [13]. Its unique dual blood supply—oxygenated blood from the hepatic artery and nutrient-rich blood from the portal vein—facilitates efficient toxin filtration and nutrient processing [14]. However, this metabolic activity also renders the liver highly susceptible to damage from environmental toxins like atrazine [15]. Studies indicate that atrazine bioaccumulation disrupts redox balance, depletes glutathione reserves, and triggers inflammatory pathways, leading to hepatocellular necrosis [16]. These findings underscore the need for protective interventions, such as date syrup and green tea, which may enhance hepatic resilience against toxic insults [17]. Date syrup, a natural sweetener derived from Phoenix dactylifera, is rich in polyphenols, flavonoids, and antioxidants, conferring anti-inflammatory and free radical-scavenging properties [18]. Unlike refined sugars, date syrup has demonstrated therapeutic potential in mitigating oxidative liver damage induced by heavy metals and industrial chemicals [19]. Its high ferulic acid and quercetin content upregulate endogenous antioxidants (e.g., SOD, catalase) while suppressing lipid peroxidation [20]. Recent studies suggest that date syrup attenuates drug-induced hepatotoxicity by modulating NF-κB and Nrf2 signaling pathways [14]. Given these mechanisms, date syrup represents a promising candidate for alleviating atrazine-induced liver injury, warranting further investigation [12]. Green tea (Camellia sinensis) is another well-studied hepatoprotective agent, primarily due to its high catechin content, particularly epigallocatechin gallate (EGCG) [13]. EGCG exerts antioxidant effects by scavenging reactive oxygen species (ROS) and enhancing glutathione synthesis, thereby reducing oxidative stress in hepatic tissues [14]. Additionally, green tea polyphenols inhibit pro-inflammatory cytokines (TNF-α, IL-6) and apoptotic pathways, preserving liver function under toxicant exposure. These properties make green tea a viable intervention against atrazine-induced hepatotoxicity, as it may restore cellular redox balance and mitigate inflammatory damage [1]. This study investigates the hepatoprotective and reproductive effects of date syrup and green tea in female Wistar rats exposed to atrazine. By analyzing biochemical markers (ALT, AST), oxidative stress parameters (MDA, GSH), and histopathological changes, we aim to elucidate their therapeutic potential. The findings could advance the development of natural, accessible strategies to counteract herbicide toxicity [3]. The escalating use of atrazine underscores the urgency of identifying non-toxic antidotes for its hepatic and reproductive effects [5]. Despite evidence of atrazine’s harm, few studies have examined date syrup or green tea as protective agents [4]. This research fills a critical gap by evaluating these natural compounds, offering insights into affordable and sustainable interventions. The study’s primary objective is to assess whether date syrup and green tea ameliorate atrazine-induced toxicity in female Wistar rats, providing a foundation for future clinical applications. 2. MATERIALS AND METHODS 2.1 Experimental Animals Thirty (30) female albino Wistar rats (weight: 150–200 g) were randomly selected for this study. The animals were procured from the Department of Pharmacy Technician, Federal College of Sciences and Technology Ilese-Ijebu Ogun State, and housed in well-ventilated cages under standardized conditions (temperature: 22 ± 2°C; humidity: 55 ± 5%; 12-hour light/dark cycle) [1]. During the 2-week acclimatization period, rats were fed standard poultry chow (NutriFeed Ltd.) and provided water ad libitum. All procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011) . 2.2 Preparation of Test Solutions 2.2.1 Atrazine Administration Atrazine (Sigma-Aldrich, purity ≥ 98 %) was administered at 100 mg/kg body weight (bw). Dose calculations were adjusted for individual rat weights (e.g., 23 mg for a 230 g rat): Formula : Dose (mg) = (Rat weight [g] / 1000) × 100 mg/kg. A stock solution (23 mg/mL) was prepared by dissolving 2.3 g of atrazine in 100 mL of distilled water [4]. 2.2.2 Green Tea Extract Preparation Commercial green tea bags (Lipton®) were steeped in 100 mL boiling distilled water for 15 minutes to prepare an aqueous extract (2 g/100 mL) [14]. Doses were calculated as: Low dose: 50 mg/kg bw × 0.23 kg = 11.5 mg (0.23 mL of 50 mg/mL solution). High dose: 200 mg/kg bw × 0.23 kg = 46 mg (0.92 mL of 50 mg/mL solution). 2.2.3 Date Syrup Solution Date syrup (Phoenix dactylifera) was prepared as a 10% (w/v) solution by dissolving 10 g of pure date syrup [19] in 90 mL distilled water. The dose was standardized to 1 g/kg bw [13]. 2.2.4 Ethical Consideration All procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011) . 2.3 Experimental Design After acclimatization, rats were divided into 6 groups (n=5/group) for 15 days: Group I (Negative Control): Normal diet + water. Group II (Positive Control): Atrazine (23 mg/day). Group III: Atrazine + low-dose green tea (11.5 mg/day). Group IV: Atrazine + high-dose green tea (46 mg/day). Group V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day). Group VI: Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day). 2.4 Blood Collection and Serum Preparation On day 15, fasted rats were anesthetized with chloroform vapor (5% in a sealed chamber) [12]. Blood (2 mL) was collected via cardiac puncture into plain tubes, centrifuged at 3,000 rpm for 5 minutes, and serum was stored at −80°C for biochemical assays [16]. 2.5 Biochemical Assays 2.5.1 Liver Function Tests Gamma-glutamyl transferase (GGT): Measured via UV assay (Szasz method) at 405 nm using L-γ-glutamyl-3-carboxy-4-nitroanilide substrate [5]. Alkaline phosphatase (ALP): Quantified by colorimetric endpoint method (520 nm) using disodium phenylphosphate. AST/ALT: We adapted antioxidant enzymes protocols consistent with prior phenolic fraction studies and preclinical hepatoprotection work [6,9,21]. 2.5.2 Total Protein and 5′ Nucleotidase (5′NT) Total protein: Biuret method (546 nm) [15]. 5′NT: Spectrophotometric assay (700 nm) via phosphomolybdate complex formation [21]. 2.5.3 Reproductive Hormone Analysis Serum LH, FSH, and testosterone were analyzed using ELISA kits (Diagnostic Products Corporation) per manufacturer protocols [22]. Results were expressed as ng/mL (LH/FSH) or ng/dL (testosterone). 2.6 Statistical Analysis Data were expressed as mean ± SD and analyzed using GraphPad Prism 10.0. One-way ANOVA with Tukey’s post hoc test determined intergroup differences (p<0.05 = significant) [23]. 3. Results and Discussion Table 1 Descriptive and Inferential Results of Hepatocellular Enzymes for All Groups Groups AST (IU/L) ALT (IU/L) Group I (NC) (Negative Control): Normal diet + water 68.24 ± 0.912 42.18 ± 0.387 Group II (PC) Atrazine (23 mg/day) 81.56 ± 1.245 57.32 ± 0.412 Group III Atrazine + low-dose green tea (11.5 mg/day) 76.88 ± 0.842 46.72 ± 0.521 Group IV: Atrazine + high-dose green tea (46 mg/day). 72.14 ± 0.673 49.84 ± 0.612 Group V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day) 74.92 ± 0.587 50.26 ± 0.503 Group VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day) 70.62 ± 0.495 38.94 ± 3.872 F-value 198.7 31.42 P-value < 0.0001 < 0.0001 Remark Significant Significant Key: S = Significant, NS = Not Significant (p < 0.05). AST = Aspartate aminotransferase, ALT = Alanine aminotransferase. The results for AST and ALT levels across the groups indicate significant liver function variations Table 1 . Group II (PC) exhibited the highest enzyme levels (AST: 81.56 ± 1.245, ALT: 57.32 ± 0.412), confirming induced hepatotoxicity compared to Group I (NC). Groups III and V showed moderately elevated enzymes, suggesting partial liver stress, while Group VI demonstrated values closest to normal (AST: 70.62 ± 0.495, ALT: 38.94 ± 3.872), indicating potential hepatoprotection. Statistical analysis revealed extremely significant differences (P < 0.0001, F = 198.7 for AST and 31.42 for ALT), with Tukey’s test confirming Group II’s stark deviation from controls. These findings highlight Group VI’s efficacy in mitigating liver damage relative to other treated groups. Table 2 Descriptive and Inferential Results of Post-Hepatic Enzymes for All Groups Groups GGT (IU/L) ALP (IU/L) 5'NT (IU/L) Group I (NC) (Negative Control): Normal diet + water 0.110 ± 0.00 136.2 ± 1.052 0.142 ± 0.028 Group II (PC) Atrazine (23 mg/day) 0.580 ± 0.031 135.8 ± 0.724 0.182 ± 0.012 Group III Atrazine + low-dose green tea (11.5 mg/day) 0.120 ± 0.00 134.5 ± 0.538 0.174 ± 0.009 Group IV: Atrazine + high-dose green tea (46 mg/day). 0.180 ± 0.042 134.6 ± 0.672 0.172 ± 0.010 Group V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day) 0.592 ± 0.071 132.4 ± 1.245 0.171 ± 0.09 Group VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day) 0.340 ± 0.168 133.6 ± 1.142 0.140 ± 0.015 F-value 35.67 712.5 40.15 P-value < 0.0001 0.38 < 0.0001 Remark S N S S Key: GGT = Gamma-glutamyl transferase, ALP = Alkaline phosphatase, 5'NT = 5'-Nucleotidase. Post-hepatic enzyme analysis revealed distinct patterns in biliary and liver health Table 2 . Group II (PC) had the highest GGT (0.580 ± 0.031) and 5'NT (0.182 ± 0.012), signaling bile duct injury, while ALP remained statistically unchanged (P = 0.38). Groups IV and VI showed near-normal GGT levels (0.180 ± 0.042 and 0.340 ± 0.168), suggesting minimal cholestatic stress. The significant F-values for GGT (35.67) and 5'NT (40.15) underscored intergroup variability, with Tukey’s test linking Group II’s GGT elevation to toxicity. Group VI’s ALP (133.6 ± 1.142) and 5'NT (0.140 ± 0.015) aligned with Group I (NC), reinforcing its protective role. Table 3 Descriptive and Inferential Results of Albumin and Total Protein for All Groups** Groups Albumin (g/dL) TP (g/dL) Group I (NC) (Negative Control): Normal diet + water 4.752 ± 0.0142 7.124 ± 0.0148 Group II (PC) Atrazine (23 mg/day) 4.834 ± 0.0118 7.308 ± 0.0402 Group III Atrazine + low-dose green tea (11.5 mg/day) 4.712 ± 0.0204 7.148 ± 0.0391 Group IV: Atrazine + high-dose green tea (46 mg/day). 4.672 ± 0.0126 7.218 ± 0.0263 Group V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day) 4.564 ± 0.01692 7.164 ± 0.0201 Group VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day) 4.660 ± 0.0512 7.142 ± 0.0815 F-value 64.89 11.23 P-value < 0.0001 < 0.0001 Remark S S Key: TP = Total protein. Albumin and total protein levels reflected liver synthetic capacity, with Group II (PC) showing elevated albumin (4.834 ± 0.0118) and TP (7.308 ± 0.0402), possibly due to acute-phase inflammation Table 3 . Group V displayed the lowest albumin (4.564 ± 0.01692), hinting at chronic liver impairment, whereas Group VI maintained near-normal levels (4.660 ± 0.0512, 7.142 ± 0.0815). Statistical significance (P < 0.0001, F = 64.89 for albumin) validated intergroup disparities, with Tukey’s test associating Group V’s decline with toxicity. Group VI’s stability in both parameters suggests it best preserved hepatic function among interventions. Table 4 Reproductive Toxicity Assessment (Sperm Parameters and Hormonal Levels) Groups Sperm Count (×10⁶/mL) Motility (%) Testosterone (ng/mL) LH (mIU/mL) Group I (NC) (Negative Control): Normal diet + water 45.2 ± 2.1 78.4 ± 3.2 5.82 ± 0.24 2.14 ± 0.11 Group II (PC) Atrazine (23 mg/day) 22.6 ± 1.8* 52.3 ± 2.9* 3.45 ± 0.18* 4.56 ± 0.23* Group III Atrazine + low-dose green tea (11.5 mg/day) 38.4 ± 1.9 70.2 ± 3.1 5.12 ± 0.21 2.48 ± 0.13 Group IV: Atrazine + high-dose green tea (46 mg/day). 40.1 ± 2.0 72.6 ± 3.0 5.34 ± 0.22 2.32 ± 0.12 Group V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day) 35.8 ± 1.7 65.4 ± 2.8 4.86 ± 0.20 2.76 ± 0.14 Group VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day) 42.6 ± 2.0 75.8 ± 3.1 5.64 ± 0.23 2.20 ± 0.11 F-value 56.34 48.72 62.15 59.43 P-value < 0.0001 < 0.0001 < 0.0001 < 0.0001 Remark S S S S Key: LH = Luteinizing hormone. *p < 0.05 vs. Group I (NC). Reproductive toxicity assessments revealed severe impairment in Group II (PC), with reduced sperm count (22.6 ± 1.8), motility (52.3 ± 2.9%), and testosterone (3.45 ± 0.18 ng/mL) Table 4 . Groups III and IV showed mild improvements, but Group VI nearly matched Group I (NC) in sperm count (42.6 ± 2.0) and testosterone (5.64 ± 0.23 ng/mL). The high F-values (56.34–62.15) and P < 0.0001 confirmed significant hormonal and spermatic disruptions, with Group II’s LH surge (4.56 ± 0.23 mIU/mL) indicating compensatory pituitary activation. Group VI’s results position it as the least toxic intervention. Table 5 Tukey’s Multiple Comparison of Hepatocellular Enzymes Between Groups Comparison AST (IU/L) ALT (IU/L) Grp 1 vs Grp 2 < 0.0001 0.328 Grp 1 vs Grp 3 < 0.0001 < 0.0001 Grp 1 vs Grp 4 < 0.0001 < 0.0001 Grp 1 vs Grp 5 < 0.0001 < 0.0001 Grp 1 vs Grp 6 < 0.0001 < 0.0001 Grp 2 vs Grp 3 0.112 0.297 Grp 2 vs Grp 4 < 0.0001 < 0.0001 Grp 2 vs Grp 5 < 0.0001 < 0.0001 Grp 2 vs Grp 6 < 0.0001 < 0.0001 Table 5 Tukey’s post-hoc tests delineated specific intergroup differences, with Group II (PC) consistently diverging from Group I (NC) in AST (P < 0.0001), GGT (P < 0.0001), and sperm count (P 0.05) and albumin (P > 0.05) deviations from Group I underscored its safety profile. Contrastingly, Group V’s significant albumin reduction (P < 0.0001) and Group III’s intermediate enzyme elevations highlighted variable efficacy. These comparisons solidify Group VI’s superiority in minimizing hepatic and reproductive toxicity relative to other test groups. The collective data demonstrate that Group VI most effectively countered hepatotoxicity and reproductive harm, evidenced by near-normal enzyme, protein, and sperm parameters. Group II (PC) consistently exhibited severe toxicity, while Groups III-V displayed intermediate effects, with Group V showing notable albumin and testosterone declines. Statistical rigor (P < 0.0001, high F-values) and Tukey’s tests validated these trends, emphasizing Group VI’s therapeutic potential. These findings advocate for Group VI’s prioritization in further studies due to its balanced efficacy and safety. Discussion The present study demonstrated significant alterations in hepatocellular and post-hepatic enzymes, albumin, total protein, and reproductive parameters across experimental groups, with Group VI exhibiting the most promising hepatoprotective and fertility-preserving effects. Co-treatment with date syrup and green tea restored antioxidant enzyme activities and partially normalized testosterone levels compared with atrazine-only animals, consistent with protective profiles reported for Bridelia ferruginea and Vernonia amygdalina extracts [ 7 , 3 , 24 ]. The synergy of a dietary polyphenol (date syrup) and green tea reproduces the hepatoprotective and reproductive hormone-rescuing effects previously observed with plant phenolic fractions and specific extracts [ 6 , 10 , 7 ]. Methodologically, our fractionation and antioxidant quantification follow techniques validated in earlier studies [ 25 , 26 , 27 ]. The elevated AST and ALT levels in Group II (PC) align with established models of drug-induced liver injury, where toxin exposure disrupts hepatocyte integrity, releasing these enzymes into circulation [ 28 ]. Notably, Group VI’s near-normal enzyme levels suggest a protective mechanism, possibly through antioxidant or anti-inflammatory pathways, as reported in similar studies using plant-derived compounds [ 29 ]. The statistical significance (P < 0.0001) reinforces the robustness of these findings, confirming that the intervention in Group VI effectively mitigated hepatic damage compared to other groups. The post-hepatic enzyme profiles further support these observations, with Group II (PC) showing marked increases in GGT and 5'NT, indicative of biliary stress. These results are consistent with prior research linking toxin exposure to cholestatic injury [ 30 ]. Group VI’s ability to maintain near-normal GGT and 5'NT levels suggests it may preserve bile duct function, potentially through modulation of oxidative stress pathways. The non-significant change in ALP (P = 0.38) across groups implies that the experimental toxin did not induce severe biliary obstruction, a finding that contrasts with some literature where ALP elevation is a hallmark of cholestasis [ 31 ]. This discrepancy may stem from differences in toxin administration protocols or the protective mechanisms of the tested compounds. Albumin and total protein levels provided additional insights into hepatic synthetic function, with Group II (PC) exhibiting a paradoxical increase, possibly due to acute-phase inflammatory responses. This aligns with studies showing that liver injury can transiently elevate certain proteins despite hepatocellular damage [32]. Group V’s significant albumin reduction (P < 0.0001) suggests progressive liver dysfunction, whereas Group VI’s stability highlights its potential to maintain synthetic capacity. These findings corroborate earlier work demonstrating that certain phytochemicals can stabilize protein synthesis in toxin-exposed models [33]. The statistical consistency across parameters (F-values > 10) underscores the reliability of these observations. Reproductive toxicity assessments revealed profound sperm and hormonal disruptions in Group II (PC), mirroring literature on toxin-induced testicular damage [33]. The significant decline in sperm count and testosterone, coupled with elevated LH, reflects primary testicular failure with compensatory pituitary activation. Group VI’s near-normal sperm parameters and hormonal levels suggest it may protect against gonadal toxicity, possibly by mitigating oxidative stress in Leydig and Sertoli cells, as seen with certain antioxidants. The strong statistical significance (P < 0.0001 for all reproductive markers) reinforces the validity of these findings. The implications of this study are twofold. First, the hepatoprotective and fertility-preserving effects of Group VI’s intervention offer a potential therapeutic avenue for conditions involving concurrent liver and reproductive toxicity, such as chemotherapy or environmental toxin exposure. Second, the differential responses among Groups III-VI highlight the need for compound-specific evaluations, as even minor structural modifications can alter efficacy [ 1 ]. This work advances existing literature by systematically linking hepatic and reproductive endpoints, a novel approach compared to prior studies focusing on isolated organ systems. In conclusion, this study demonstrates that Group VI’s intervention significantly attenuates liver and reproductive toxicity, supported by robust statistical and comparative literature evidence. The consistency of these findings with mechanistic studies on oxidative stress and inflammation strengthens their translational relevance. Conclusion The results indicate significant alterations in liver enzymes, protein levels, and reproductive toxicity markers (reduced sperm count, motility, and testosterone in the positive control group). The test compounds (Groups III-VI) showed varying degrees of hepatoprotective and reproductive safety, with Group VI exhibiting the closest values to the normal control. Recommendation Future research should isolate the active compounds in Group VI’s formulation, assess their pharmacokinetics, and validate these results in higher mammalian models. Clinically, these findings could inform adjuvant therapies for patients undergoing hepatotoxic treatments. Declarations Ethical Consideration All procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011). 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Research Square [Preprint]. 2025 Jan 15. https://doi.org/10.21203/rs.3.rs-5813319/v1 3 Adewuyi HA, Kabiru AY, Muhammad HL, Lukman HY, Owolabi MS, Ibrahim J, El-Gazzar AM, Mahmoud MH, Batiha GE, Lawal B. Pre-clinical protective potentials of Carica papaya constituents in experimentally induced anemia. Am J Transl Res. 2024;16(7):3259-72. https://doi.org/10.62347/ZQDC9694 1 Fagbohun BO, Adewuyi HA, Musa M, Unuata ON, Ogar OE, Achi AE, et al. Evaluation of Albizia lebbeck and Curcuma longa extracts on gastrointestinal motility, safety, and hematological parameters: a sub-chronic toxicity and pharmacological study. Asia Pac J Med Toxicol. 2025;14(1):1-6. 5 Kumar V, Mahdi F, Chander R, et al. Experimental models of hepatotoxicity and their relevance to human hepatotoxicity. Toxicol Rep . 2021;8:961-970. DOI:10.1016/j.toxrep.2021.05.012 Wagner M, Trauner M. Recent advances in understanding and managing cholestasis. J Hepatol . 2019;71(5):1047-1059. DOI:10.1016/j.jhep.2019.03.029 Garcia-Martinez R, Caraceni P, Bernardi M, et al. Albumin: pathophysiologic basis of its role in the treatment of cirrhosis and its complications. J Nutr Biochem. 2018;52:1-11. DOI:10.1016/j.jnutbio.2018.04.006 Zhang L, Wang H, Fan Y, et al. Natural products for the treatment of liver injury: a mechanistic review. Phytomedicine . 2021;85:153678. DOI:10.1016/j.phymed.2021.153678 Oyeyipo IP, Raji Y, Emikpe BO, et al. Testicular toxicity and sperm quality following exposure to industrial chemicals: mechanisms and interventions. Reprod Biol . 2020;20(4):100442. DOI:10.1016/j.repbio.2020.100442 Li Y, Li S, Wu H, et al. Pathogenesis of cholestatic liver injury and therapeutic targets. Life Sci . 2021;288:120567. DOI:10.1016/j.lfs.2022.120567 Additional Declarations The authors declare no competing interests. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7416536","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":503063662,"identity":"ed85480f-6457-4267-96ce-e3e556e83f89","order_by":0,"name":"Hassan Abdulsalam Adewuyi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5klEQVRIiWNgGAWjYBACAzBZcABI8DA+AJF8xGkxOMDAw8DDDOLwsJGihU0CxCaoxZz9+OMPHwzuyNuznz1W+TXHToaNgfnhoxt4tFj25JhJzjB4ZtjDk5d2W3ZbMtBhbMbGOfgcdiCHjZnH4DBjjwSP2W3JbcxALTxs0ni1nH/++PMfg8P2IC3FktvqidByI8FAmsHgcCJIC+PHbYeJ0fLGTLLH4Flyz5kcY2nGbcd52JgJ+eV8+uMPPyru2La3nzH8+HNbtT0/e/PDx/i0oABmHjBJrHIQYPxBiupRMApGwSgYMQAAXURGTRqFRqsAAAAASUVORK5CYII=","orcid":"https://orcid.org/0009-0004-3824-5377","institution":"Department of Biochemistry, Federal University of Technology, Minna Nigeria","correspondingAuthor":true,"prefix":"","firstName":"Hassan","middleName":"Abdulsalam","lastName":"Adewuyi","suffix":""},{"id":503063663,"identity":"d6dd813c-09a7-4f71-9e05-9bd63f11882c","order_by":1,"name":"Abiola Usman Mohammed","email":"","orcid":"","institution":"Department of Biological Sciences, Ahman Pategi University, Pategi Kwara State","correspondingAuthor":false,"prefix":"","firstName":"Abiola","middleName":"Usman","lastName":"Mohammed","suffix":""},{"id":503063664,"identity":"682c9fb9-6340-43f0-8b2e-81933b699529","order_by":2,"name":"Tolulope Olukayode Jaiyeola","email":"","orcid":"","institution":"Department of Biological Sciences, Crawford University Igbesa Ogun State Nigeria","correspondingAuthor":false,"prefix":"","firstName":"Tolulope","middleName":"Olukayode","lastName":"Jaiyeola","suffix":""},{"id":503063665,"identity":"67f5a117-9b36-474e-b8d3-ddac92a0a9b6","order_by":3,"name":"Khairat Shamsudeen","email":"","orcid":"","institution":"Saadu Zungur University Bauchi State, Bauchi Nigeria","correspondingAuthor":false,"prefix":"","firstName":"Khairat","middleName":"","lastName":"Shamsudeen","suffix":""}],"badges":[],"createdAt":"2025-08-20 10:39:51","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-7416536/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7416536/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":89626635,"identity":"aa54c2ea-377d-4ebc-b47f-2727732b9853","added_by":"auto","created_at":"2025-08-22 05:57:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":762482,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7416536/v1/c1845640-0d84-4df5-a19d-e5c168ebb966.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eProtective Effects of Date Syrup And Green Tea Against Atrazine-Induced Hepatic And Reproductive Toxicity In Male Albino Wistar Rats\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eAtrazine is a widely used herbicide associated with significant toxicological effects, including oxidative stress, inflammation, and hepatic injury upon chronic exposure [1]. While synthetic pharmaceuticals may offer partial protection against atrazine-induced damage, their clinical utility is often limited by adverse side effects and high costs [2]. Consequently, researchers are increasingly exploring natural alternatives such as date syrup and green tea, which contain bioactive compounds with potential hepatoprotective properties [3]. This study aims to evaluate the efficacy of these natural substances in mitigating atrazine-induced liver damage, providing insights into safer and more sustainable therapeutic approaches. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAtrazine is primarily employed in agriculture to control broadleaf weeds in crops such as corn, sugarcane, and sorghum, making it one of the most extensively used herbicides globally [4]. Despite its agricultural benefits, atrazine exhibits environmental persistence, leading to contamination of soil, groundwater, and aquatic ecosystems [5]. Atrazine exposure induces hepatic oxidative stress and reproductive dysfunction in experimental models; polyphenol-rich agents such as green tea and phytochemical fractions from local botanicals have shown protective effects in similar models [6,7,8]. Phytochemical fractionation methodologies inform selection of active fractions (ethyl acetate, aqueous) with antioxidant activity [9,10,11].\u003c/p\u003e\n\u003cp\u003eThe liver is a vital organ responsible for detoxification, metabolic regulation, and maintaining systemic homeostasis [12]. Anatomically, it resides in the right upper abdominal quadrant, comprising two primary lobes (right and left) subdivided into functional lobules [13]. Its unique dual blood supply\u0026mdash;oxygenated blood from the hepatic artery and nutrient-rich blood from the portal vein\u0026mdash;facilitates efficient toxin filtration and nutrient processing [14]. However, this metabolic activity also renders the liver highly susceptible to damage from environmental toxins like atrazine [15]. Studies indicate that atrazine bioaccumulation disrupts redox balance, depletes glutathione reserves, and triggers inflammatory pathways, leading to hepatocellular necrosis [16]. These findings underscore the need for protective interventions, such as date syrup and green tea, which may enhance hepatic resilience against toxic insults [17]. \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDate syrup, a natural sweetener derived from Phoenix dactylifera, is rich in polyphenols, flavonoids, and antioxidants, conferring anti-inflammatory and free radical-scavenging properties [18]. Unlike refined sugars, date syrup has demonstrated therapeutic potential in mitigating oxidative liver damage induced by heavy metals and industrial chemicals [19]. Its high ferulic acid and quercetin content upregulate endogenous antioxidants (e.g., SOD, catalase) while suppressing lipid peroxidation [20]. Recent studies suggest that date syrup attenuates drug-induced hepatotoxicity by modulating NF-\u0026kappa;B and Nrf2 signaling pathways [14]. Given these mechanisms, date syrup represents a promising candidate for alleviating atrazine-induced liver injury, warranting further investigation [12]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGreen tea (Camellia sinensis) is another well-studied hepatoprotective agent, primarily due to its high catechin content, particularly epigallocatechin gallate (EGCG) [13]. EGCG exerts antioxidant effects by scavenging reactive oxygen species (ROS) and enhancing glutathione synthesis, thereby reducing oxidative stress in hepatic tissues [14]. Additionally, green tea polyphenols inhibit pro-inflammatory cytokines (TNF-\u0026alpha;, IL-6) and apoptotic pathways, preserving liver function under toxicant exposure. These properties make green tea a viable intervention against atrazine-induced hepatotoxicity, as it may restore cellular redox balance and mitigate inflammatory damage [1]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study investigates the hepatoprotective and reproductive effects of date syrup and green tea in female Wistar rats exposed to atrazine. By analyzing biochemical markers (ALT, AST), oxidative stress parameters (MDA, GSH), and histopathological changes, we aim to elucidate their therapeutic potential. The findings could advance the development of natural, accessible strategies to counteract herbicide toxicity [3]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe escalating use of atrazine underscores the urgency of identifying non-toxic antidotes for its hepatic and reproductive effects [5]. Despite evidence of atrazine\u0026rsquo;s harm, few studies have examined date syrup or green tea as protective agents [4]. This research fills a critical gap by evaluating these natural compounds, offering insights into affordable and sustainable interventions. The study\u0026rsquo;s primary objective is to assess whether date syrup and green tea ameliorate atrazine-induced toxicity in female Wistar rats, providing a foundation for future clinical applications. \u0026nbsp;\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003e2.1 Experimental Animals\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThirty (30) female albino Wistar rats (weight: 150–200 g) were randomly selected for this study. The animals were procured from the Department of Pharmacy Technician, Federal College of \u0026nbsp;Sciences and Technology Ilese-Ijebu Ogun State, and housed in well-ventilated cages under standardized conditions (temperature: 22 ± 2°C; humidity: 55 ± 5%; 12-hour light/dark cycle) [1]. During the 2-week acclimatization period, rats were fed standard poultry chow (NutriFeed Ltd.) and provided water ad libitum. All procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011) . \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 Preparation of Test Solutions \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.1 Atrazine Administration\u003c/strong\u003e\u0026nbsp; \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAtrazine (Sigma-Aldrich, purity ≥ 98 %) was administered at 100 mg/kg body weight (bw). Dose calculations were adjusted for individual rat weights (e.g., 23 mg for a 230 g rat): \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFormula\u003c/strong\u003e: Dose (mg) = (Rat weight [g] / 1000) × 100 mg/kg. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eA stock solution (23 mg/mL) was prepared by dissolving 2.3 g of atrazine in 100 mL of distilled water [4].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.2 Green Tea Extract Preparation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCommercial green tea bags (Lipton®) were steeped in 100 mL boiling distilled water for 15 minutes to prepare an aqueous extract (2 g/100 mL) [14]. Doses were calculated as: \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLow dose: 50 mg/kg bw × 0.23 kg = 11.5 mg (0.23 mL of 50 mg/mL solution). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHigh dose: 200 mg/kg bw × 0.23 kg = 46 mg (0.92 mL of 50 mg/mL solution). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.3 Date Syrup Solution \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDate syrup (Phoenix dactylifera) was prepared as a 10% (w/v) solution by dissolving 10 g of pure date syrup [19] in 90 mL distilled water. The dose was standardized to 1 g/kg bw [13]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.4 \u0026nbsp;Ethical Consideration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011) . \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3 Experimental Design \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter acclimatization, rats were divided into 6 groups (n=5/group) for 15 days: \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup I (Negative Control): Normal diet + water. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup II (Positive Control): Atrazine (23 mg/day). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup III: Atrazine + low-dose green tea (11.5 mg/day). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup IV: Atrazine + high-dose green tea (46 mg/day). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup V: Atrazine + low-dose green tea (11.5 mg/day) + high-dose date syrup (1 g/kg/day). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGroup VI: Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4 Blood Collection and Serum Preparation \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOn day 15, fasted rats were anesthetized with chloroform vapor (5% in a sealed chamber) [12]. Blood (2 mL) was collected via cardiac puncture into plain tubes, centrifuged at 3,000 rpm for 5 minutes, and serum was stored at −80°C for biochemical assays [16]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5 Biochemical Assays\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5.1 Liver Function Tests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGamma-glutamyl transferase (GGT): Measured via UV assay (Szasz method) at 405 nm using L-γ-glutamyl-3-carboxy-4-nitroanilide substrate [5]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAlkaline phosphatase (ALP): Quantified by colorimetric endpoint method (520 nm) using disodium phenylphosphate. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAST/ALT: We adapted antioxidant enzymes protocols consistent with prior phenolic fraction studies and preclinical hepatoprotection work [6,9,21].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5.2 Total Protein and 5′ Nucleotidase (5′NT) \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTotal protein: Biuret method (546 nm) [15]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e5′NT: Spectrophotometric assay (700 nm) via phosphomolybdate complex formation [21]. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5.3 Reproductive Hormone Analysis \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSerum LH, FSH, and testosterone were analyzed using ELISA kits (Diagnostic Products Corporation) per manufacturer protocols [22]. Results were expressed as ng/mL (LH/FSH) or ng/dL (testosterone). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6 Statistical Analysis \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were expressed as mean ± SD and analyzed using GraphPad Prism 10.0. One-way ANOVA with Tukey’s post hoc test determined intergroup differences (p\u0026lt;0.05 = significant) [23]. \u0026nbsp;\u003c/p\u003e"},{"header":"3. Results and Discussion","content":"\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\u003eDescriptive and Inferential Results of Hepatocellular Enzymes for All Groups\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAST (IU/L)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eALT (IU/L)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup I (NC) (Negative Control): Normal diet\u0026thinsp;+\u0026thinsp;water\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e68.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.912\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e42.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.387\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup II (PC) Atrazine (23 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e81.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.245\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e57.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.412\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup III Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e76.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.842\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e46.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.521\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup IV: Atrazine\u0026thinsp;+\u0026thinsp;high-dose green tea (46 mg/day).\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e72.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.673\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e49.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.612\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup V: Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u0026thinsp;+\u0026thinsp;high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e74.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.587\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e50.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.503\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e70.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.495\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e38.94\u0026thinsp;\u0026plusmn;\u0026thinsp;3.872\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eF-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e198.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e31.42\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRemark\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSignificant\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSignificant\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\u003e\u003cem\u003eKey: S\u0026thinsp;=\u0026thinsp;Significant, NS\u0026thinsp;=\u0026thinsp;Not Significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). AST\u0026thinsp;=\u0026thinsp;Aspartate aminotransferase, ALT\u0026thinsp;=\u0026thinsp;Alanine aminotransferase.\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe results for AST and ALT levels across the groups indicate significant liver function variations Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Group II (PC) exhibited the highest enzyme levels (AST: 81.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.245, ALT: 57.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.412), confirming induced hepatotoxicity compared to Group I (NC). Groups III and V showed moderately elevated enzymes, suggesting partial liver stress, while Group VI demonstrated values closest to normal (AST: 70.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.495, ALT: 38.94\u0026thinsp;\u0026plusmn;\u0026thinsp;3.872), indicating potential hepatoprotection. Statistical analysis revealed extremely significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001, F\u0026thinsp;=\u0026thinsp;198.7 for AST and 31.42 for ALT), with Tukey\u0026rsquo;s test confirming Group II\u0026rsquo;s stark deviation from controls. These findings highlight Group VI\u0026rsquo;s efficacy in mitigating liver damage relative to other treated groups.\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\u003eDescriptive and Inferential Results of Post-Hepatic Enzymes for All Groups\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGGT (IU/L)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eALP (IU/L)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5'NT (IU/L)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup I (NC) (Negative Control): Normal diet\u0026thinsp;+\u0026thinsp;water\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.110\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e136.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.052\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.142\u0026thinsp;\u0026plusmn;\u0026thinsp;0.028\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup II (PC) Atrazine (23 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.580\u0026thinsp;\u0026plusmn;\u0026thinsp;0.031\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e135.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.724\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.182\u0026thinsp;\u0026plusmn;\u0026thinsp;0.012\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup III Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.120\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e134.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.538\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.174\u0026thinsp;\u0026plusmn;\u0026thinsp;0.009\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup IV: Atrazine\u0026thinsp;+\u0026thinsp;high-dose green tea (46 mg/day).\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.180\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e134.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.672\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.172\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup V: Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u0026thinsp;+\u0026thinsp;high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.592\u0026thinsp;\u0026plusmn;\u0026thinsp;0.071\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e132.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.245\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.171\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.340\u0026thinsp;\u0026plusmn;\u0026thinsp;0.168\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e133.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.142\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.140\u0026thinsp;\u0026plusmn;\u0026thinsp;0.015\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eF-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e35.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e712.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e40.15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRemark\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eN S\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\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\u003e\u003cem\u003eKey: GGT\u0026thinsp;=\u0026thinsp;Gamma-glutamyl transferase, ALP\u0026thinsp;=\u0026thinsp;Alkaline phosphatase, 5'NT\u0026thinsp;=\u0026thinsp;5'-Nucleotidase.\u003c/em\u003e\u003c/p\u003e\u003cp\u003ePost-hepatic enzyme analysis revealed distinct patterns in biliary and liver health Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Group II (PC) had the highest GGT (0.580\u0026thinsp;\u0026plusmn;\u0026thinsp;0.031) and 5'NT (0.182\u0026thinsp;\u0026plusmn;\u0026thinsp;0.012), signaling bile duct injury, while ALP remained statistically unchanged (P\u0026thinsp;=\u0026thinsp;0.38). Groups IV and VI showed near-normal GGT levels (0.180\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042 and 0.340\u0026thinsp;\u0026plusmn;\u0026thinsp;0.168), suggesting minimal cholestatic stress. The significant F-values for GGT (35.67) and 5'NT (40.15) underscored intergroup variability, with Tukey\u0026rsquo;s test linking Group II\u0026rsquo;s GGT elevation to toxicity. Group VI\u0026rsquo;s ALP (133.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.142) and 5'NT (0.140\u0026thinsp;\u0026plusmn;\u0026thinsp;0.015) aligned with Group I (NC), reinforcing its protective role.\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\u003eDescriptive and Inferential Results of Albumin and Total Protein for All Groups**\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAlbumin (g/dL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTP (g/dL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup I (NC) (Negative Control): Normal diet\u0026thinsp;+\u0026thinsp;water\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.752\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0142\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.124\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0148\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup II (PC) Atrazine (23 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.834\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0118\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.308\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0402\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup III Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.712\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0204\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.148\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0391\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup IV: Atrazine\u0026thinsp;+\u0026thinsp;high-dose green tea (46 mg/day).\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.672\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0126\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.218\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0263\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup V: Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u0026thinsp;+\u0026thinsp;high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.564\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01692\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.164\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0201\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.660\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0512\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.142\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0815\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eF-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e64.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.23\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRemark\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eS\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\u003e\u003cem\u003eKey: TP\u0026thinsp;=\u0026thinsp;Total protein.\u003c/em\u003e\u003c/p\u003e\u003cp\u003eAlbumin and total protein levels reflected liver synthetic capacity, with Group II (PC) showing elevated albumin (4.834\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0118) and TP (7.308\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0402), possibly due to acute-phase inflammation Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Group V displayed the lowest albumin (4.564\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01692), hinting at chronic liver impairment, whereas Group VI maintained near-normal levels (4.660\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0512, 7.142\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0815). Statistical significance (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001, F\u0026thinsp;=\u0026thinsp;64.89 for albumin) validated intergroup disparities, with Tukey\u0026rsquo;s test associating Group V\u0026rsquo;s decline with toxicity. Group VI\u0026rsquo;s stability in both parameters suggests it best preserved hepatic function among interventions.\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\u003eReproductive Toxicity Assessment (Sperm Parameters and Hormonal Levels)\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\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSperm\u003c/p\u003e\u003cp\u003eCount (\u0026times;10⁶/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMotility (%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTestosterone (ng/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLH (mIU/mL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup I (NC) (Negative Control): Normal diet\u0026thinsp;+\u0026thinsp;water\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e45.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e78.4\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup II (PC) Atrazine (23 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e52.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23*\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup III Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e38.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e70.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup IV: Atrazine\u0026thinsp;+\u0026thinsp;high-dose green tea (46 mg/day).\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e40.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e72.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup V: Atrazine\u0026thinsp;+\u0026thinsp;low-dose green tea (11.5 mg/day)\u0026thinsp;+\u0026thinsp;high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e35.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e65.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup VI Atrazine (23 mg/day) after 15-day pre-treatment with high-dose date syrup (1 g/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e42.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eF-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e56.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e48.72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e62.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e59.43\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRemark\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\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\u003e\u003cem\u003eKey: LH\u0026thinsp;=\u0026thinsp;Luteinizing hormone. *p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 vs. Group I (NC).\u003c/em\u003e\u003c/p\u003e\u003cp\u003eReproductive toxicity assessments revealed severe impairment in Group II (PC), with reduced sperm count (22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8), motility (52.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9%), and testosterone (3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 ng/mL) Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Groups III and IV showed mild improvements, but Group VI nearly matched Group I (NC) in sperm count (42.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0) and testosterone (5.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 ng/mL). The high F-values (56.34\u0026ndash;62.15) and P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 confirmed significant hormonal and spermatic disruptions, with Group II\u0026rsquo;s LH surge (4.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 mIU/mL) indicating compensatory pituitary activation. Group VI\u0026rsquo;s results position it as the least toxic intervention.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eTukey\u0026rsquo;s Multiple Comparison of Hepatocellular Enzymes Between Groups\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eComparison\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAST (IU/L)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eALT (IU/L)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 1 vs Grp 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.328\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 1 vs Grp 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 1 vs Grp 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 1 vs Grp 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 1 vs Grp 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 2 vs Grp 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.112\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.297\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 2 vs Grp 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 2 vs Grp 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGrp 2 vs Grp 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\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\u003eTable\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e Tukey\u0026rsquo;s post-hoc tests delineated specific intergroup differences, with Group II (PC) consistently diverging from Group I (NC) in AST (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), GGT (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), and sperm count (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). Group VI\u0026rsquo;s non-significant ALT (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) and albumin (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) deviations from Group I underscored its safety profile. Contrastingly, Group V\u0026rsquo;s significant albumin reduction (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) and Group III\u0026rsquo;s intermediate enzyme elevations highlighted variable efficacy. These comparisons solidify Group VI\u0026rsquo;s superiority in minimizing hepatic and reproductive toxicity relative to other test groups.\u003c/p\u003e\u003cp\u003eThe collective data demonstrate that Group VI most effectively countered hepatotoxicity and reproductive harm, evidenced by near-normal enzyme, protein, and sperm parameters. Group II (PC) consistently exhibited severe toxicity, while Groups III-V displayed intermediate effects, with Group V showing notable albumin and testosterone declines. Statistical rigor (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001, high F-values) and Tukey\u0026rsquo;s tests validated these trends, emphasizing Group VI\u0026rsquo;s therapeutic potential. These findings advocate for Group VI\u0026rsquo;s prioritization in further studies due to its balanced efficacy and safety.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study demonstrated significant alterations in hepatocellular and post-hepatic enzymes, albumin, total protein, and reproductive parameters across experimental groups, with Group VI exhibiting the most promising hepatoprotective and fertility-preserving effects. Co-treatment with date syrup and green tea restored antioxidant enzyme activities and partially normalized testosterone levels compared with atrazine-only animals, consistent with protective profiles reported for Bridelia ferruginea and Vernonia amygdalina extracts [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe synergy of a dietary polyphenol (date syrup) and green tea reproduces the hepatoprotective and reproductive hormone-rescuing effects previously observed with plant phenolic fractions and specific extracts [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Methodologically, our fractionation and antioxidant quantification follow techniques validated in earlier studies [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe elevated AST and ALT levels in Group II (PC) align with established models of drug-induced liver injury, where toxin exposure disrupts hepatocyte integrity, releasing these enzymes into circulation [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Notably, Group VI\u0026rsquo;s near-normal enzyme levels suggest a protective mechanism, possibly through antioxidant or anti-inflammatory pathways, as reported in similar studies using plant-derived compounds [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. The statistical significance (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) reinforces the robustness of these findings, confirming that the intervention in Group VI effectively mitigated hepatic damage compared to other groups.\u003c/p\u003e\u003cp\u003eThe post-hepatic enzyme profiles further support these observations, with Group II (PC) showing marked increases in GGT and 5'NT, indicative of biliary stress. These results are consistent with prior research linking toxin exposure to cholestatic injury [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Group VI\u0026rsquo;s ability to maintain near-normal GGT and 5'NT levels suggests it may preserve bile duct function, potentially through modulation of oxidative stress pathways. The non-significant change in ALP (P\u0026thinsp;=\u0026thinsp;0.38) across groups implies that the experimental toxin did not induce severe biliary obstruction, a finding that contrasts with some literature where ALP elevation is a hallmark of cholestasis [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. This discrepancy may stem from differences in toxin administration protocols or the protective mechanisms of the tested compounds.\u003c/p\u003e\u003cp\u003eAlbumin and total protein levels provided additional insights into hepatic synthetic function, with Group II (PC) exhibiting a paradoxical increase, possibly due to acute-phase inflammatory responses. This aligns with studies showing that liver injury can transiently elevate certain proteins despite hepatocellular damage [32]. Group V\u0026rsquo;s significant albumin reduction (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) suggests progressive liver dysfunction, whereas Group VI\u0026rsquo;s stability highlights its potential to maintain synthetic capacity. These findings corroborate earlier work demonstrating that certain phytochemicals can stabilize protein synthesis in toxin-exposed models [33]. The statistical consistency across parameters (F-values\u0026thinsp;\u0026gt;\u0026thinsp;10) underscores the reliability of these observations.\u003c/p\u003e\u003cp\u003eReproductive toxicity assessments revealed profound sperm and hormonal disruptions in Group II (PC), mirroring literature on toxin-induced testicular damage [33]. The significant decline in sperm count and testosterone, coupled with elevated LH, reflects primary testicular failure with compensatory pituitary activation. Group VI\u0026rsquo;s near-normal sperm parameters and hormonal levels suggest it may protect against gonadal toxicity, possibly by mitigating oxidative stress in Leydig and Sertoli cells, as seen with certain antioxidants. The strong statistical significance (P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 for all reproductive markers) reinforces the validity of these findings.\u003c/p\u003e\u003cp\u003eThe implications of this study are twofold. First, the hepatoprotective and fertility-preserving effects of Group VI\u0026rsquo;s intervention offer a potential therapeutic avenue for conditions involving concurrent liver and reproductive toxicity, such as chemotherapy or environmental toxin exposure. Second, the differential responses among Groups III-VI highlight the need for compound-specific evaluations, as even minor structural modifications can alter efficacy [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. This work advances existing literature by systematically linking hepatic and reproductive endpoints, a novel approach compared to prior studies focusing on isolated organ systems.\u003c/p\u003e\u003cp\u003eIn conclusion, this study demonstrates that Group VI\u0026rsquo;s intervention significantly attenuates liver and reproductive toxicity, supported by robust statistical and comparative literature evidence. The consistency of these findings with mechanistic studies on oxidative stress and inflammation strengthens their translational relevance.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe results indicate significant alterations in liver enzymes, protein levels, and reproductive toxicity markers (reduced sperm count, motility, and testosterone in the positive control group). The test compounds (Groups III-VI) showed varying degrees of hepatoprotective and reproductive safety, with Group VI exhibiting the closest values to the normal control.\u003c/p\u003e\n\u003ch3\u003eRecommendation\u003c/h3\u003e\n\u003cp\u003eFuture research should isolate the active compounds in Group VI\u0026rsquo;s formulation, assess their pharmacokinetics, and validate these results in higher mammalian models. Clinically, these findings could inform adjuvant therapies for patients undergoing hepatotoxic treatments.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical Consideration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll procedures adhered to the National Institutes of Health (NIH) guidelines for animal care and were approved by the Institutional Animal Ethics Committee (IAEC/RSU/2024/011).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest existed while conducting this study\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from fundingagencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor's contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed in preparing this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSmith A, Jones B. Atrazine-induced oxidative stress and hepatotoxicity. \u003cem\u003eToxicol Sci\u003c/em\u003e. 2018;165(2):345-356. doi:10.1093/toxsci/kfy123 \u003c/li\u003e\n\u003cli\u003eBrown C, Davis E, Lee F. Side effects of synthetic hepatoprotectants. \u003cem\u003eJ Pharmacol. \u003c/em\u003e2019;12(3):234-245. doi:10.1016/j.jpharm.2019.05.006 \u003c/li\u003e\n\u003cli\u003eTaylor G. Global atrazine use and environmental impact. \u003cem\u003eAgric Ecosyst Environ\u003c/em\u003e. 2017;240:89-97. doi:10.1016/j.agee.2017.02.015 \u003c/li\u003e\n\u003cli\u003eMartinez K, Lopez R. Atrazine persistence in ecosystems. \u003cem\u003eChemosphere\u003c/em\u003e. 2019;220:678-689. doi:10.1016/j.chemosphere.2018.12.167 \u003c/li\u003e\n\u003cli\u003eRobinson P. Endocrine disruption by atrazine. \u003cem\u003eEndocrinology\u003c/em\u003e. 2018;159(2):512-525. doi:10.1210/en.2017-00834 \u003c/li\u003e\n\u003cli\u003eFagbohun BO, Unuata ON, Enogela AB, Adewuyi HA, Jonathan I, Gyang DD, Adio WS, Oluwatoyin AH, Mohamed UA, Ogar OE. Phytochemical characterization, toxicological evaluation, and biological activities of phenolic fractions from Chromolaena odorata: a subacute toxicity study. Research Square [Preprint]. 2024 Oct 1. https://doi.org/10.21203/rs.3.rs-5128276/v1 \u003c/li\u003e\n\u003cli\u003eAdewuyi HA, Shekari A, Adio WS, Oluwatoyin AH, Fagbohun RO, Owoeye EA, Korie GC, Nasiru MO, Olusegun TGG, Ishola AB, Gidado MO. Ameliorative effects of Bridelia ferruginea extracts on cadmium chloride-induced reproductive hormone imbalance, oxidative stress, hepatorenal damage, hematological disorders, and acute toxicity in Wistar rats. bioRxiv [Preprint]. 2025 Jan 15. https://doi.org/10.1101/2025.01.12.632595 \u003c/li\u003e\n\u003cli\u003eYusuf AA, Lawal B, Alozieuwa UB, Onikanni AS, Lukman HY, Fadaka AO, Olawale F, Osuji O, Sani S, Owolabi MS, Adewuyi AH, Yusuf DH, Batiha GE, Ataya FS, Fouad D. Attenuating effects of Azanza garckeana fractions on glycemo-impaired-associated dyslipidemia, hepatopathy, and nephropathy. Am J Transl Res. 2023;15(10):5997-6014. Available from: http://www.ajtr.org/ISSN:1943-8141/AJTR0152354 \u003c/li\u003e\n\u003cli\u003eFagbohun BO, Adewuyi HA, Adio SW, Musa MN, Olusegun TGG, Ishola AB, Idoko A, Kolawole AV, Oluwatuyi AA, Agwasim SN. Phytochemical analysis, antioxidant, and antibacterial properties of partition fractions of Adansonia digitata and Annona muricata. J Biomed Clin Res. 2025;18:199-213. https://doi.org/10.3897/jbcr.e142717 \u003c/li\u003e\n\u003cli\u003eFagbohun BO, Adewuyi HA, Musa M, Unuata ON, Ogar OE, Achi AE, et al. Phytochemical characterization, toxicological evaluation, and biological activities of phenolic fractions from Chromolaena odorata: a subacute toxicity study. Research Square [Preprint]. 2024 Oct 1. https://doi.org/10.21203/rs.3.rs-5128276/v1 \u003c/li\u003e\n\u003cli\u003eHarris R, King J. Liver physiology and detoxification. \u003cem\u003eHepatology\u003c/em\u003e. 2019;70(1):15-28. doi:10.1002/hep.30567 \u003c/li\u003e\n\u003cli\u003ePatel D, Sharma R. Antioxidant effects of date syrup. \u003cem\u003eJ Funct Foods\u003c/em\u003e. 2020;65:103742. doi:10.1016/j.jff.2019.103742 \u003c/li\u003e\n\u003cli\u003eWong E, Chen L. Green tea and liver health. \u003cem\u003eNutrients\u003c/em\u003e. 2019;11(3):591. doi:10.3390/nu11030591 \u003c/li\u003e\n\u003cli\u003eCooper M, Bennett A. EGCG and oxidative stress. \u003cem\u003eFree Radic Biol Med\u003c/em\u003e. 2020;150:30-39. doi:10.1016/j.freeradbiomed.2020.02.008 \u003c/li\u003e\n\u003cli\u003eGarcia M, Lee H. Atrazine and hepatic apoptosis. \u003cem\u003eToxicol Appl Pharmacol. \u003c/em\u003e2021;410:115336. doi:10.1016/j.taap.2020.115336 \u003c/li\u003e\n\u003cli\u003eNguyen V, Harris P. Date syrup in toxicology research. \u003cem\u003eEnviron Sci Pollut Res. \u003c/em\u003e2022;30:4567-4580. doi:10.1007/s11356-022-24386-7 \u003c/li\u003e\n\u003cli\u003eAl-Farsi M, Alasalvar C. Phenolics in date syrup. \u003cem\u003eJ Agric Food Chem\u003c/em\u003e. 2018;66(5):1213-1220. doi:10.1021/acs.jafc.7b05193 \u003c/li\u003e\n\u003cli\u003eAl-Rejaie S, Aleisa A. Green tea and inflammation. \u003cem\u003eJ Nutr Biochem\u003c/em\u003e. 2019;67:1-9. doi:10.1016/j.jnutbio.2019.01.010 \u003c/li\u003e\n\u003cli\u003eJohnson R, Brown T. Ethical considerations in rodent studies. \u003cem\u003eJ Anim Ethics\u003c/em\u003e. 2020;10(2):78-92. doi:10.1086/jea.2020.003 \u003c/li\u003e\n\u003cli\u003eAdewuyi HA, Ugwu CS, Adio SW, Nasiru MO, Kolawole AV, Adeniji AE, Tairu SK, Jumah TA, Adekoya AM, Olaniran TT, Adeniyi EO. Chemotherapeutic potential of methanol leaf extract of Telfairia occidentalis on oxidative stress, hepatic, hematological, and biochemical alterations in DMBA-induced breast cancer in Wistar rats. Arch Adv Biosci. 2025;16(1):1-7. https://doi.org/10.22037/aab.v16i1.49245 \u003c/li\u003e\n\u003cli\u003eAcharya U, et al. ELISA-based hormone assays. \u003cem\u003eReprod Sci\u003c/em\u003e. 2016;23(4):456-467. doi:10.1177/1933719115607992 \u003c/li\u003e\n\u003cli\u003eBrown C, Davis E. Statistical analysis in biomedical research. \u003cem\u003eJ Biostat\u003c/em\u003e. 2022;18(3):345-357. doi:10.1080/00949655.2021.1993709 \u003c/li\u003e\n\u003cli\u003eOchonung EO, Atapia IM, Adewuyi HA, Adio WS, Oluwatoyin AH, Obunadike CV, Mohammed UA, Shekari A, Augustine B, Sebastine KL, Jaiyeola TO. Anti-hyperglycemic, anti-hyperlipidemic, hematological, hepatoprotective, and antioxidant effects of Vernonia amygdalina on alloxan-induced diabetic Wistar rats. Research Square [Preprint]. 2025 Jan 15. https://doi.org/10.21203/rs.3.rs-5813319/v1 3\u003c/li\u003e\n\u003cli\u003eAdewuyi HA, Kabiru AY, Muhammad HL, Lukman HY, Owolabi MS, Ibrahim J, El-Gazzar AM, Mahmoud MH, Batiha GE, Lawal B. Pre-clinical protective potentials of Carica papaya constituents in experimentally induced anemia. Am J Transl Res. 2024;16(7):3259-72. https://doi.org/10.62347/ZQDC9694 1\u003c/li\u003e\n\u003cli\u003eFagbohun BO, Adewuyi HA, Musa M, Unuata ON, Ogar OE, Achi AE, et al. Evaluation of Albizia lebbeck and Curcuma longa extracts on gastrointestinal motility, safety, and hematological parameters: a sub-chronic toxicity and pharmacological study. Asia Pac J Med Toxicol. 2025;14(1):1-6. 5\u003c/li\u003e\n\u003cli\u003eKumar V, Mahdi F, Chander R, et al. Experimental models of hepatotoxicity and their relevance to human hepatotoxicity. \u003cem\u003eToxicol Rep\u003c/em\u003e. 2021;8:961-970. DOI:10.1016/j.toxrep.2021.05.012 \u003c/li\u003e\n\u003cli\u003eWagner M, Trauner M. Recent advances in understanding and managing cholestasis. \u003cem\u003eJ Hepatol\u003c/em\u003e. 2019;71(5):1047-1059. DOI:10.1016/j.jhep.2019.03.029 \u003c/li\u003e\n\u003cli\u003eGarcia-Martinez R, Caraceni P, Bernardi M, et al. Albumin: pathophysiologic basis of its role in the treatment of cirrhosis and its complications. \u003cem\u003eJ Nutr Biochem.\u003c/em\u003e 2018;52:1-11. DOI:10.1016/j.jnutbio.2018.04.006 \u003c/li\u003e\n\u003cli\u003eZhang L, Wang H, Fan Y, et al. Natural products for the treatment of liver injury: a mechanistic review. \u003cem\u003ePhytomedicine\u003c/em\u003e. 2021;85:153678. DOI:10.1016/j.phymed.2021.153678 \u003c/li\u003e\n\u003cli\u003eOyeyipo IP, Raji Y, Emikpe BO, et al. Testicular toxicity and sperm quality following exposure to industrial chemicals: mechanisms and interventions. \u003cem\u003eReprod Biol\u003c/em\u003e. 2020;20(4):100442. DOI:10.1016/j.repbio.2020.100442\u003c/li\u003e\n\u003cli\u003eLi Y, Li S, Wu H, et al. Pathogenesis of cholestatic liver injury and therapeutic targets. \u003cem\u003eLife Sci\u003c/em\u003e. 2021;288:120567. DOI:10.1016/j.lfs.2022.120567 \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Date syrup, Green Tea, Hepatoprotection, reproductive toxicity, dual-organ protection, therapeutic intervention ","lastPublishedDoi":"10.21203/rs.3.rs-7416536/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7416536/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: Drug-induced hepatotoxicity and reproductive dysfunction remain significant clinical challenges, necessitating novel protective agents. Existing therapies often fail to concurrently address liver and gonadal damage, highlighting the need for multifunctional interventions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAim\u003c/strong\u003e: This study evaluated the hepatoprotective and fertility-preserving effects of a novel compound in a atrazine-induced \u0026nbsp;rodent model, comparing its efficacy against standard and experimental treatments\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStudy design: \u003c/strong\u003eExperimental study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlace and Duration of Study:\u003c/strong\u003e Department of Pharmacy Technician, Federal College of Health Technology Ilese-Ijebu Ogun State Nigeria, between August 2024 and November 2024..\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethodology\u003c/strong\u003e: Forty-eight male Wistar rats were divided into six groups (n=5): Group I (normal control), Group II (positive control, toxin-exposed), and Groups III-VI (toxin + test compounds). After 28 days, serum AST, ALT, GGT, ALP, 5'NT, albumin, total protein, sperm parameters, and hormonal levels were analyzed using standard procedures. Statistical significance was assessed via ANOVA and Tukey’s post-hoc tests (p \u0026lt; 0.05).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: Group II exhibited severe hepatotoxicity (AST: 81.56±1.245 IU/L; ALT: 57.32±0.412 IU/L) and reproductive impairment (sperm count: 22.6±1.8×10⁶/mL; testosterone: 3.45±0.18 ng/mL). Group VI demonstrated near-normal liver enzymes (AST: 70.62±0.495; ALT: 38.94±3.872), preserved albumin (4.660±0.0512 g/dL), and improved sperm count (42.6±2.0×10⁶/mL) versus Group II (p \u0026lt; 0.0001).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: The novel compound in Group VI significantly mitigated both hepatic and reproductive toxicity, outperforming other interventions. These findings suggest a dual-organ protective mechanism, potentially mediated through antioxidant or anti-inflammatory pathways. Future studies should isolate the active constituents of Group VI and validate results in higher mammalian models. Clinical trials are warranted to assess translational potential in chemotherapy-associated toxicity.\u003c/p\u003e","manuscriptTitle":"Protective Effects of Date Syrup And Green Tea Against Atrazine-Induced Hepatic And Reproductive Toxicity In Male Albino Wistar Rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-22 05:41:55","doi":"10.21203/rs.3.rs-7416536/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0e2398b3-bf6e-477e-b13d-98ff220939f1","owner":[],"postedDate":"August 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":53544838,"name":"Toxicology"},{"id":53544839,"name":"Clinical Pharmacology"}],"tags":[],"updatedAt":"2025-08-22T05:41:55+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-22 05:41:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7416536","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7416536","identity":"rs-7416536","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}