Impact of Vitamin D3 Supplementation on Motor Functionality and the Immune Response in Parkinson's Disease Patients with Vitamin D Deficiency

Impact of Vitamin D3 Supplementation on Motor Functionality and the Immune Response in Parkinson's Disease Patients with Vitamin D Deficiency | 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 Impact of Vitamin D3 Supplementation on Motor Functionality and the Immune Response in Parkinson's Disease Patients with Vitamin D Deficiency Danfeng LI, Xibo Ma, Wentao Zhang, Ping Zhong, Xiaowei Zhu, Yan Chen, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5145248/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 The purpose of this study was to investigate the levels of vitamin D, regulatory T (Treg) cells, and T helper 17 (Th17) cells in the peripheral blood of patients with Parkinson's disease(PD), explore their potential relationships with PD, and further examine the effects of vitamin D intervention on these biomarkers and clinical outcomes. PD patients with low vitamin D3 levels were randomly assigned to two groups: those supplemented with vitamin D3 (VitD3, n=15) and those treated with vegetable oil (PL, n=15). Treatment was administered continuously for 3 months. Vitamin D3, Th17 and Treg levels and related clinical scales were continuously monitored and evaluated. The results revealed that the level of 25(OH) D3 in the peripheral blood increased in the VitD3 group, and the Th17 and Treg levels tended toward those of the normal population. Compared with those of the preintervention groups and the PL group after the 3-month follow-up, motor function improved in the VitD3 group. Additionally, vitamin D was negatively correlated with Th17 cells and positively correlated with Treg cells. In PD patients, motor function was positively correlated with vitamin D and Treg levels but negatively correlated with Th17 levels. We believe that it is necessary for PD patients with low vitamin D levels to supplement vitamin D to normal levels. Parkinson's disease Th17 Treg vitamin D Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Parkinson's disease, the second most prevalent neurodegenerative disorder, affects approximately 1–2% of individuals aged 65 and older. Projections suggest that by 2040, the global prevalence of PD will exceed 12 million individuals worldwide. However, the exact etiology of PD remains unclear, with multiple factors implicated 1 . The intricate mechanism of neurodegeneration in PD has yet to be fully elucidated, but it is believed to encompass a complex interplay of genetic and environmental factors, oxidative stress, mitochondrial dysfunction, inflammation, immune modulation, and other factors. Vitamin D is a steroid hormone that performs many other functions in addition to its well-known role in calcium homeostasis. There is extensive evidence from in vitro and animal studies indicating that vitamin D plays crucial roles in cell proliferation and differentiation, neurotrophic regulation and neuroprotection, neurotransmission, immune regulation, and neural plasticity 2-6 . Furthermore, the expression of vitamin D receptor (VDR) and CYP27B1 is particularly abundant in the substantia nigra, a brain region rich in dopaminergic neurons 7 . As a fat-soluble hormone, vitamin D can cross the blood‒brain barrier, which serves as indirect evidence supporting its role in the central nervous system 8 . There are four main subtypes of CD4+ T cells: Th1, Th2, Th17, and Treg cells. Among them, Th17 cells are a proinflammatory subtype, whereas Treg cells are an anti-inflammatory subtype. The balance between Th17 and Treg cell levels plays a crucial role in maintaining immune homeostasis and inducing antigen-specific immune tolerance 9, 10 . Various autoimmune diseases are associated with the disruption of this balance 11 . Research over the past decade has shown that these immune alterations are not limited to the periphery but also extend to the central nervous system, where they promote the progression of neurodegenerative diseases 12 . In cellular models, Th17 cells have been demonstrated to disrupt and cross the blood‒brain barrier and are associated with increased neuroinflammation in central nervous system pathologies 13, 14 . Within the central nervous system, regulatory T cells actively promote neural system recovery through the inhibition of astrocyte proliferation, with particular emphasis on their role in ischemic stroke and neuroinflammatory diseases 15 . Similar results have also been reported in studies involving PD patients, where Th17 cells were found to induce the apoptosis of midbrain neurons 16 , whereas Tregs exhibited a protective effect on dopamine neurons 17 . Many studies have reported increased proportions of Th17 cells and decreased proportions and impaired functions of Treg cells in patients with Parkinson's disease 18, 19 . Vitamin D can regulate Th17 and Treg cells primarily by inducing the expression of the Foxp3 gene in naive CD4+ T cells 20, 21 . The VDR binds to VDREs within the Foxp3 gene, enabling vitamin D to directly upregulate Foxp3 expression, thereby promoting the differentiation and expression of Treg cells. Conversely, Foxp3 inhibits the expression of RORγt in CD4+T cells 22 . RORγt is the primary transcription regulator of Th17 cells, promoting and maintaining their specificity as Th17 cells. Therefore, inhibiting RORγt expression leads to the suppression of Th17 differentiation and expression. In summary, vitamin D regulates the balance between Th17 and Treg cells, playing a key role in maintaining immune homeostasis. Low vitamin Dlevels in PD patients have been recognized for more than 15 years 23 , and numerous clinical studies have corroborated the imbalance of Th17 and Treg levels in these patients 18, 19, 24 . We hypothesized that it is possible to regulate the homeostatic balance between Th17 and Treg cells by improving low vitamin D levels in Parkinson's disease patients, thereby improving clinical symptoms. This study aimed to detect the expression levels of vitamin D, Tregs, and Th17 cells in the peripheral blood of PD patients, explore the influence of Treg/Th17 imbalance on PD progression, and further investigate the impact of vitamin D intervention on this imbalance and clinical outcomes. We hope to gain a deeper understanding of the role of vitamin D deficiency and the Treg/Th17 imbalance in PD. 2. Materials And Methods 2.1. Design of the Study This study was a randomized, double-blind, placebo-controlled clinical trial (NCT: 06539260) that administered a three-month supply of vitamin D 3 and a placebo(vegetable oil) to patients with PD who had low levels of vitamin D. This study obtained approval from the Ethics Committee of Suzhou Hospital of Anhui Medical University, with approval number A2023024. The procedures used in this study adhered to the principles of the Declaration of Helsinki. All patients signed informed consent to participate in the study and were informed of their right to withdraw at any point during the trial. 2.2. Participants From January 2023 to March 2024, 51 PD patients were recruited. All the subjects were from the Parkinson's outpatient clinic and inpatient ward of the Neurology Department at Suzhou Hospital of Anhui Medical University. Fifty partners or volunteers of PD patients, matched by sex and age, were selected from Suzhou Hospital of Anhui Medical University to form a healthy control(HC) group. The inclusion criteria for patients were as follows: (1) met the diagnostic criteria for PD established by the UK Parkinson's Disease Society Brain Bank. (2) Age between 45 and 80 years. The exclusion criteria were as follows: (1) Having related vitamin D metabolic diseases (such as renal failure, severe liver damage, inherited 1α-hydroxylase deficiency, etc.). (2) Having immune system diseases. (3) Having a history of disabling cerebrovascular disease. (4) Having first- or second-degree relatives with PD. (5) Having severe dementia, depression, or severe mental illness. 2.3. Randomization and Masking For the PD patients included in the study, analyses were conducted on their peripheral blood vitamin D levels. PD patients with low vitamin D levels were divided into two groups at a 1:1 ratio to receive vitamin D 3 and PL. Randomization was performed via an Excel random number generator. Both vitamin D 3 and the PL were placed in identical bottles labeled only with numbers and containing no additional information. Product allocation and randomization were carried out by an independent researcher. Both the personnel involved in data analysis and the patients remained blinded until the database was examined for analysis. Unblinding occurred when all the data were analyzed. 2.4. Outcome The primary endpoint of this study focused on the changes observed from baseline (T0) to the 3-month mark (T1). We evaluated PD patients via a comprehensive set of scales, including the Movement Disorder Society Unified Parkinson's Disease Rating Scale (MDS-UPDRS), Berg Balance Scale (Breg), Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE), Self-rating Anxiety Scale (SAS), Self-rating Depression Scale (SDS), and Parkinson's Disease Sleep Scale (PDSS). During the initial T0 stage, we analyzed the peripheral blood levels of Th17, Treg, and Vit D in the PD patients and healthy volunteers. Among the PD patients with low vitamin D levels, we randomly assigned them into two groups: one to receive vitamin D 3 supplementation and the other to receive PL. At the T1 stage, we followed up with both groups of PD patients to reassess the indicators and monitor any changes (Fig. 1). 2.5. Measurement of vitamin D The serum 25(OH)D 3 concentration was measured via the Elecsys method on a Roche electrochemiluminescence fully automated immunoassay analyzer. According to the guidelines of the Endocrine Society, normal vitamin D levels are defined as ≥30 ng/mL, 20–30 ng/mL is defined as insufficient, and vitamin deficiency is defined as ≤20 ng/mL 25 . 2.6. Measurement of Th17 and Treg Venous blood withdrawal was performed after a night of fasting, between 8:00 a.m. and 10:00 a.m., into heparin lithium anticoagulant tubes. The tubes were subsequently coded and stored at room temperature until processing, which occurred 12 hours after collection, to ensure homogeneous treatment of all the samples. Th17 cells by flow cytometry The blood samples were diluted 1:1 with high-sugar medium (RPMI 1640) and mixed well. Leukocyte activation cocktail (3 μL/1 ml, BD Pharmingen) was added to the solution of diluted blood. The mixture was incubated for 4–6 hours in 5% CO2 at 37°C. Two milliliters of hemolysin (BD Pharmingen) was added, and the mixture was vortexed and incubated for 10 minutes at room temperature in the dark. The samples were subsequently centrifuged, the supernatants were removed, the cells were washed in PBS, and the resulting mixture was resuspended in PBS. One hundred microliters of PBS buffer was added, the cells were resuspended, FITC-conjugated mouse anti-human CD3 (BD Pharmingen) and CD8-conjugated PERCP-CY5.5 (BioLegend) were added for surface antibody staining, and the mixture was incubated for 15 minutes in the dark. After the cells were fixed and permeabilized (Fixation/Permeabilization Kit, BD Pharmingen), an intracellular PE-conjugated mouse anti-human IL-17A antibody (BD Pharmingen) was added, and the mixture was incubated at room temperature in the dark for 30 minutes. Assessment of Tregs by flow cytometry CD4-FITC and CD25-PE antibodies (BD Pharmingen) were added to the flow tube. Then, 50 µl of peripheral blood was added to the tube, which was vortexed and mixed well. The mixture was incubated in the dark for 15 minutes. Then, 500 μL of hemolysin (BD Pharmingen) was added, the mixture was shaken, and the mixture was incubated in the dark for 15 minutes. Three milliliters of PBS was added, the mixture was centrifuged for 5 minutes, and the supernatant was removed. The cells were fixed and permeabilized (FOXP3/TRN FACTOR STAIN BUFFER, eBioscience). The intranuclear staining antibody foxp3 (ANTI-HU FOXP3 PCH101 APC, eBioscience) was added to each tube, and the mixture was incubated at 4°C for 2 hours in the dark. Three milliliters of PBS was added again, the mixture was centrifuged for 5 minutes, and the supernatant was removed. Acquisition was then performed on a BD FACSCanto II flow cytometer. CD3 + CD8 - IL-17 + cells were identified as Th17 cells, whereas CD4+CD25+Foxp3+ cells were identified as Treg cells. Lymphocytes are recognized on the basis of their classical forward scatter (FSC) and side scatter (SSC) signals. The data were analyzed via FlowJo software (version 10.8.1). 2.7 Statistical analysis In this study, SPSS 26.0 and GraphPad Prism v.9 software were used for data analysis. Descriptive statistical methods were employed to analyze all the data gathered during the research process. For intergroup comparisons, an independent sample t test was conducted, a paired sample t test was used for comparisons before and after the follow-up. Pearson correlation analysis was applied to data adhering to a normal distribution, whereas Spearman correlation analysis was employed for data deviating from a normal distribution. A P value of less than 0.05 was considered statistically significant. 3. Results 3.1 Demographic characteristics This study included a total of 51 PD patients and 50 healthy individuals, all of whom were of Chinese ethnicity and fulfilled the established inclusion criteria. The PD group included 29 males and 22 females, with a mean age of 69.6±11.0 years. The HCs included 26 males and 24 females, with a mean age of 65.3±10.9 years. No statistically significant differences were observed in sex composition or age between the two groups (P>0.05). The PD group presented a mean H-Y grade of 2.32±0.90, an average UPDRS score of 45.76±23.06, a mean UPDRS-III score of 26.35±13.56, an average Breg score of 43.45±12.18, a mean MMSE score of 21.46±7.07, an average MoCA score of 18.32±7.51, a mean SDS self-rating depression scale score of 45.85±13.47, a mean SAS self-rating anxiety scale score of 44.20±12.50, and an average PDSS score of 100.82±23.48, as indicated in Table 1. The PD patients enrolled were categorized into a normal vitamin D level group (Vit D≥30 ng/ml), consisting of 21 patients, and a low vitamin D level group (Vit D<30 ng/ml), comprising 30 patients, on the basis of their baseline vitamin D levels. The low vitamin D group was subsequently randomly divided into two subgroups, a VitD 3 group and a PL group, each comprising 15 patients. Importantly, no complications arose during or following vitamin D supplementation for any of the subjects with vitamin D deficiency. 3.2 Comparison of 25(OH) D 3 , Th17, and Treg cells in the PD Group and HC Group To investigate changes in CD4+ T-cell subsets in the peripheral blood, we first evaluated the percentages of CD4+ T-cell subsets (Th17 and Treg) in 51 PD patients and 50 healthy controls. Compared with the HCs, the PD cohort presented significant differences in Th17 levels (3.78±1.33 vs 2.30±0.79, t=6.799, p<0.001; Fig. 2a) and Treg levels (4.16±1.29 vs 4.64±0.97, t=-2.142, p=0.035; Fig. 2b). Additionally, there was a significant difference in the serum 25(OH) D 3 level between the PD cohort and the HC cohort (28.98±8.10 vs 35.80±6.96, t=-4.531; p<0.001; Fig. 2c). 3.3 Vitamin D, Th17, and Treg correlations in human blood We conducted a correlation analysis on a total of 101 data sets from the PD group and the HC group. The analysis revealed a positive correlation between vitamin D levels in peripheral blood and the percentage of Tregs (r=0.526, p<0.001; Fig. 3a) and a negative correlation with the percentage of Th17 cells (r=-0.635, p<0.001; Fig. 3b). 3.4 Correlations between PD-related scores and Th17, Treg, and vitamin D levels The proportion of Th17 cells was positively correlated with both the UPDRS and the UPDRS-III score (r=0.412, p=0.003 and r=0.432, p=0.002, respectively) and negatively correlated with the Breg balance scale (r=-0.302, P=0.031). The proportion of Treg cells was negatively correlated with the UPDRS and UPDRS-III scores (r=-0.504, p<0.001 and r=-0.540, p<0.001, respectively) and positively correlated with the Breg balance scale (r=0.382, p=0.006). A correlation analysis between vitamin D levels and UPDRS and UPDRS-III scores among PD patients revealed a negative correlation (r=-0.494, p<0.001 and r=-0.549, p<0.001, respectively) and a positive correlation with Breg (r=0.419, p=0.002), as indicated in Table 2. These results indicate that the severity of motor impairment in Parkinson's disease patients is correlated with the levels of Th17, Treg, and vitamin D. The MMSE score of PD patients is positively correlated with Th17 levels but negatively correlated with vitamin D levels. The MoCA score is positively correlated with Th17 levels and negatively correlated with Treg and vitamin D levels. On the other hand, no correlation was observed between the SDS and SAS scores of PD patients and the levels of Th17, Treg, and vitamin D. Additionally, the PDSS score positively correlated with Treg but was not correlated with Th17 and vitamin D, as indicated in Table 2. Therefore, the intellectual status of PD patients seems to be associated with the levels of Th17, Treg, and vitamin D. 3.5 Follow-up results There were 30 patients with low vitamin D levels in the PD group; these patients were randomly divided into the VitD 3 subgroup 15 patients and the PL subgroup 15 patients. The basic characteristics of the two subgroups are shown in Table 3. After a three-month follow-up with medication administration, the VitD 3 subgroup exhibited a notable reduction in Th17 levels during vitamin D supplementation, with levels decreasing from 4.62±1.09 to 3.25±1.14 (P=0.003, Fig. 4a). In contrast, Th17 levels did not significantly change in the PL subgroup before or after the three-month period (p>0.05, Fig. 4a). The Treg levels in the VitD 3 subgroup increased (3.25±0.90 vs 4.52±0.95, P=0.003; Fig. 4b), whereas there was no significant difference in the Treg levels before and after treatment in the PL subgroup (p>0.05; Fig. 4b). Additionally, we evaluated the VitD 3 subgroup of PD patients via the UPDRS, UPDRS-III, and Breg scales to assess three motor indicators before and after treatment. We found that the VitD 3 subgroup showed significant improvement in motor indicators after three months of vitamin D supplementation, with UPDRS scores improving from 57.00±20.86 to 52.27±21.38 (P=0.003, Fig. 4c) and UPDRS-III scores improving from 32.40±11.70 to 28.13±12.44 (P<0.001, Fig. 4d). However, there was no difference in the Breg scale score before and after treatment (39.73±13.36 vs 40.60±12.91, P=0.400; Fig. 4e). Furthermore, there were no differences in motor indicators before and after three months in the PL subgroup, as indicated in Supplementary Table 1. 4. Discussion Our findings indicate that the average vitamin D level in PD patients is lower than that in healthy controls, which is consistent with the results of several previous clinical studies 23 . Additionally, PD patients have a higher Th17/Treg ratio than healthy individuals do. Upon comprehensive analysis, we observed a significant correlation between the levels of 25(OH) D 3 in human peripheral blood and Th17 and Treg cells. This finding is in line with our hypothesis that 25-hydroxyvitamin D 3 interacts with the vitamin D receptor on the surface of CD4+ T cells, participating in their differentiation and expression and promoting the expression and differentiation of Treg cells while inhibiting the differentiation and expression of Th17 cells. For the analysis of motor function in PD patients, we used the UPDRS-III and Breg balance scales for assessment. There was a significant correlation between the UPDRS-III and Breg scores of PD patients and the levels of Th17, Treg, and 25(OH) D 3 . Furthermore, our analysis of nonmotor scales revealed a significant correlation between the cognitive scales (MoCA, MMS) of PD patients and the levels of Th17, Treg, and 25(OH) D 3 . A three-month follow-up was conducted on both the VitD 3 group and the PL group. The results revealed that the levels of 25(OH) D 3 in the peripheral blood of the VitD 3 group were increased, and the levels of Th17 and Treg cells tended toward those of the normal population. Furthermore, compared with their preintervention levels and those of the PL group at the three-month follow-up, the motor function of the VitD 3 group had improved. The role of vitamin D in PD has been extensively studied. In some studies, vitamin D deficiency has been observed among PD patients 26 . Research has revealed a correlation between the severity of motor impairments in PD patients and their peripheral blood vitamin D levels 27, 28 , and our study yielded similar results. In 2010, a longitudinal study based on a health survey of the Finnish population revealed that individuals with low vitamin D levels had a 65% greater probability of developing Parkinson's disease than did those with high vitamin D levels 29 . These findings indirectly support the role of vitamin D in the onset and progression of PD. Vitamin D can achieve neuroprotective effects by inhibiting the early aggregation of α-synuclein (α-Syn), especially in neurodegenerative diseases 30 . A study on the treatment of Parkinson's disease in animal models with vitamin D revealed that vitamin D has neuroprotective effects by attenuating proinflammatory processes and upregulating anti-inflammatory processes in animal models of PD 31 . Vitamin D mediates the secretion and expression of inflammatory factors, which may be the pathway through which it influences the onset and progression of PD. In 1988, McGeer et al. discovered HLA-DR-reactive microglia in the postmortem substantia nigra tissue of Parkinson's disease patients 32 , which was one of the earliest lines of evidence linking neuroinflammation to the pathogenesis of Parkinson's disease. Microglia, monocytes, and infiltrating T cells, which migrate around α-syn in PD patients' substantia nigra, suggest both the role of innate and adaptive immunity in PD 33 . A recent study revealed that α-Syn levels in PD patients are associated with the severity of motor symptoms and an imbalance of Th17/Treg cells 18 . Furthermore, a previous study suggested that α-Syn impairs the stability of Tregs and promotes the differentiation of Th17 cells in PD 18 . Contact between Th17 cells and midbrain neurons can directly lead to the apoptosis of dopaminergic neurons 16, 34 , and IL-17 has a destructive effect on the blood‒brain barrier 35 . Conversely, Treg cells can significantly protect the survival of midbrain dopaminergic neurons 17 , which also indirectly indicates that an imbalance of Th17/Treg cells in PD patients may exacerbate clinical symptoms. Treg cells play a significant role in reducing the beta-amyloid protein load in Alzheimer's patients, restoring brain homeostasis, and improving learning and memory 36 . Similarly, our study revealed a clear correlation between Treg levels and cognitive function in PD patients, suggesting that improving Treg levels may be a potential therapeutic approach for PD patients with cognitive issues. α-Syn promotes the differentiation of Th17 cells and activates intracellular inflammatory pathways through autoimmune effects 18 , and low vitamin D levels may exacerbate this situation 30 . Either through the direct immunomodulatory effects of vitamin D or through the immune pathways mediated by α-syn, low vitamin D levels lead to an imbalance between Th17 cells and Treg cells 37 , causing damage to substantia nigra neurons 16, 17 . We believe that this pathway represents a promising direction for future research on the pathogenesis of Parkinson's disease and may lead to the discovery of new treatments. This study has several limitations. First, the relatively small sample size restricts the generalizability of the results to a limited scope. Second, the observed vitamin D deficiency in PD patients could stem from reduced outdoor activity and inadequate sunlight exposure due to motor disorders, subsequently hindering vitamin D synthesis. However, we have not undertaken any supplementary analysis specifically addressing this underlying mechanism. Third, while our study revealed improvements in Th17 and Treg levels, as well as motor function, among PD patients receiving vitamin D supplementation, we remain unable to definitively attribute the motor improvement solely to the restoration of neural function resulting from the improved Th17/Treg balance. In the future, further experiments are needed to explore the deeper mechanisms by which vitamin D affects the balance between Th17 and Treg cells and to verify the neuroprotective effect of vitamin D on PD patients through improvements in their autoimmune status. 5. Conclusion In the present study, we observed that, compared with healthy individuals, PD patients presented elevated levels of Th17 and decreased levels of Treg and vitamin D. The more pronounced these numerical deviations are, the more severe the motor dysfunction symptoms become in PD patients. Supplementing PD patients with low vitamin D levels could regulate their Th17 and Treg levels, reduce autoimmune inflammation, and improve motor function. In summary, we believe that restoring vitamin D levels to normal is necessary for PD patients with vitamin D deficiency, as it can increase their self-care ability and help alleviate or slow the progression of disability. Our data support the notion that vitamin D plays a crucial role in mediating the balance between Th17 and Treg cells, which is closely linked to the pathogenesis of Parkinson's disease. Declarations Acknowledgments The authors have no acknowledgments to report. Funding Statement This research was supported by the Scientific Research Project of the Health Commission of Anhui Province (AHWJ2022b106) and the Anhui Provincial Key Research and Development Plan (202204295107020063). Conflict of interest All the authors declare that this research was conducted without any commercial or financial relationships that could be construed as potential conflicts of interest. Data availability statement The data supporting the findings of this study are available within the article and its supplementary material. Ethical approval This study was conducted in accordance with ethical principles that originated in the Declaration of Helsinki and that are consistent with good clinical practice. Institutional review boards or independent ethics committees provided written approval for the study protocol and all amendments. All patients provided written informed consent. ClinicalTrials.gov: NCT:06539260. Registered 05 August 2024 - Retrospectively registered, https://clinicaltrials.gov/study/NCT06539260. Author contributions Danfeng Li and Xibo Ma wrote this manuscript. Yan Chen, Xiaowei Zhu, and Xibo Ma participated in data collection. 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Mov Disord 2012; 27: 264-271. 20111231. DOI: 10.1002/mds.24016. Ding. H, Dhima. K, Lockhart. KC, et al. Unrecognized vitamin D3 deficiency is common in Parkinson disease. Neurology 2013: 1531–1537. DOI: 10.1212/WNL.0b013e3182a95818. Knekt P, Kilkkinen A, Rissanen H, et al. Serum vitamin D and the risk of Parkinson disease. Arch Neurol 2010; 67: 808-811. DOI: 10.1001/archneurol.2010.120. Zhang Y, Ji W, Zhang S, et al. Vitamin D Inhibits the Early Aggregation of alpha-Synuclein and Modulates Exocytosis Revealed by Electrochemical Measurements. Angew Chem Int Ed Engl 2022; 61: e202111853. 20211122. DOI: 10.1002/anie.202111853. Calvello R, Cianciulli A, Nicolardi G, et al. Vitamin D Treatment Attenuates Neuroinflammation and Dopaminergic Neurodegeneration in an Animal Model of Parkinson's Disease, Shifting M1 to M2 Microglia Responses. J Neuroimmune Pharmacol 2017; 12: 327-339. 20161216. DOI: 10.1007/s11481-016-9720-7. McGeer PL, Itagaki S and BE. B. Reactive microglia are positive for HLA-DR in the substantia nigra of Parkinson’s and Alzheimer’s disease brains. Neurology 1988; 38: 1285-1291. DOI: 10.1212/wnl.38.8.1285. Tansey MG, Wallings RL, Houser MC, et al. Inflammation and immune dysfunction in Parkinson disease. Nat Rev Immunol 2022; 22: 657-673. 20220304. DOI: 10.1038/s41577-022-00684-6. Lindestam Arlehamn CS, Dhanwani R, Pham J, et al. alpha-Synuclein-specific T cell reactivity is associated with preclinical and early Parkinson's disease. Nat Commun 2020; 11: 1875. 20200420. DOI: 10.1038/s41467-020-15626-w. Liu Z, Qiu AW, Huang Y, et al. IL-17A exacerbates neuroinflammation and neurodegeneration by activating microglia in rodent models of Parkinson's disease. Brain Behav Immun 2019; 81: 630-645. 20190724. DOI: 10.1016/j.bbi.2019.07.026. Yeapuri P, Machhi J, Lu Y, et al. Amyloid-beta specific regulatory T cells attenuate Alzheimer's disease pathobiology in APP/PS1 mice. Mol Neurodegener 2023; 18: 97. 20231218. DOI: 10.1186/s13024-023-00692-7. Ao T, Kikuta J and Ishii M. The Effects of Vitamin D on Immune System and Inflammatory Diseases. Biomolecules 2021; 11 20211103. DOI: 10.3390/biom11111624. Tables Table 1 Characteristics of PD patients and healthy controls. PD Group (n=51) HC Group (n=50) Sex M: 29 W: 22 M: 26 W: 24 Age 69.6±11.0 65.3±10.9 H-Y 2.32±0.90 - UPDRS 45.76±23.06 - UPDRS-III 26.35±13.56 - Breg 43.45±12.18 - MMSE 21.46±7.07 - MoCA 18.32±7.51 - SDS 45.85±13.47 - SAS 44.20±12.50 - PDSS 100.82±23.48 - Table 2 Correlations between PD-related scores and Th17, Treg, and vitamin D levels. Th17 Treg Vitamin D r p r p r p UPDRS 0.412 0.003 -0.504 0.001 -0.494 0.001 UPDRS-III 0.432 0.002 -0.540 0.001 -0.549 0.001 Breg -0.302 0.031 0.382 0.006 0.419 0.002 MMSE -0.352 0.012 0.161 0.263 0.316 0.025 MoCA -0.377 0.007 0.308 0.029 0.374 0.007 SDS -0.012 0.933 -0.148 0.299 -0.108 0.450 SAS 0.111 0.440 -0.210 0.138 -0.224 0.114 PDSS -0.050 0.727 0.347 0.013 -0.137 0.338 Table 3 Characteristics of the VitD group and PL group. VitD (n=15) PL (n=15) p Sex M: 9 W: 6 M: 9 W: 6 P >0.05 Age 73.4±8.3 73.1±10.8 P >0.05 H-Y 2.63±0.77 2.67±0.96 P >0.05 Th17 4.62±1.09 4.41±1.15 P >0.05 Treg 3.25±0.90 3.61±0.93 P >0.05 Vitamin D 22.98±3.79 23.84±3.93 P >0.05 UPDRS 57.0±20.9 54.4±25.7 P >0.05 UPDRS-III 32.4±11.7 -34.0±14.4 P >0.05 Breg 39.7±13.4 37.3±13.8 P >0.05 Supplementary Files Picture1.jpg SupplementaryTable1.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-5145248","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":376582849,"identity":"30125236-4d94-4f59-af01-9ba34a34904d","order_by":0,"name":"Danfeng LI","email":"","orcid":"","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":false,"prefix":"","firstName":"Danfeng","middleName":"","lastName":"LI","suffix":""},{"id":376582850,"identity":"ae869b0a-9d66-4456-b3f1-5c88c104bfba","order_by":1,"name":"Xibo Ma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAsUlEQVRIiWNgGAWjYFACHsYDDAxycmzs7QeI1sIAVGpszMdzJoE0LYnzJBwMiNOgOyP3wGHeNoP0NgmGBIYfFdsIazE7cy7hMM8Zg9w26cYDjD1nbhOh5XiPwcEZFX9y22QOJDAzthGj5TAPUIuBQTqbRIIBkVqAthz4UGGQQIIWoF8OfDhjYNgGDOSDxPnlRu7BB4ltBvLy7e0HH/yoIEILCjhAovpRMApGwSgYBbgAAIu2Ps1atOKvAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3369-0066","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":true,"prefix":"","firstName":"Xibo","middleName":"","lastName":"Ma","suffix":""},{"id":376582851,"identity":"34801a64-92ec-431e-a39d-620fc98f3e11","order_by":2,"name":"Wentao Zhang","email":"","orcid":"","institution":"Tongji University Tenth People's Hospital: Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Wentao","middleName":"","lastName":"Zhang","suffix":""},{"id":376582852,"identity":"890b6a01-0da6-4a14-8b63-fcbdb9bdb693","order_by":3,"name":"Ping Zhong","email":"","orcid":"","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":false,"prefix":"","firstName":"Ping","middleName":"","lastName":"Zhong","suffix":""},{"id":376582853,"identity":"58680803-4826-4a19-adf1-8962907d7b28","order_by":4,"name":"Xiaowei Zhu","email":"","orcid":"","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xiaowei","middleName":"","lastName":"Zhu","suffix":""},{"id":376582854,"identity":"038ee52d-6c17-42a3-8d6e-5b1848df76d5","order_by":5,"name":"Yan Chen","email":"","orcid":"","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Chen","suffix":""},{"id":376582855,"identity":"99395882-d668-40c6-9268-c1a7d1d331d4","order_by":6,"name":"Shihua Liu","email":"","orcid":"","institution":"Suzhou Hospital of Anhui Medical University","correspondingAuthor":false,"prefix":"","firstName":"Shihua","middleName":"","lastName":"Liu","suffix":""}],"badges":[],"createdAt":"2024-09-24 12:59:38","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5145248/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5145248/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":71550439,"identity":"c744ad27-1405-4ebb-b280-93e8b48632f2","added_by":"auto","created_at":"2024-12-16 15:39:51","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":63589,"visible":true,"origin":"","legend":"\u003cp\u003eFlow diagram of the study\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/c06db851da68c9221747ec7d.jpg"},{"id":71550438,"identity":"47c1906c-7453-4583-a9f3-ed77245e1adc","added_by":"auto","created_at":"2024-12-16 15:39:51","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":133866,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of 25(OH) D3, Th17, and Treg between PD patients and HCs\u003c/p\u003e","description":"","filename":"Picture3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/1263bc60d553f9e142d9228f.jpg"},{"id":71550890,"identity":"2d9de5f6-7efa-4a6b-9cc5-2f893198e497","added_by":"auto","created_at":"2024-12-16 15:47:51","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":88931,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelations of vitamin D with Th17 and Treg cells\u003c/p\u003e","description":"","filename":"Picture4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/c670f9fa306825cec83de977.jpg"},{"id":71550888,"identity":"7beee2c3-478b-4dab-bc95-3b255f832662","added_by":"auto","created_at":"2024-12-16 15:47:51","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":180275,"visible":true,"origin":"","legend":"\u003cp\u003eEffects of supplementation on Th17, Treg, and motor indicators in the VitD and PL groups\u003c/p\u003e","description":"","filename":"Picture5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/5186e0065b7e9da1a3ef63ca.jpg"},{"id":72341606,"identity":"0a7a243b-7344-48a6-a8ca-62f9910483e1","added_by":"auto","created_at":"2024-12-25 18:58:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":885016,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/228c8325-57b1-49d0-bcd3-c7cd218895bc.pdf"},{"id":71550454,"identity":"22aed252-d098-4485-936e-f9e8b7cd273c","added_by":"auto","created_at":"2024-12-16 15:39:52","extension":"jpg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":130463,"visible":true,"origin":"","legend":"","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/ec4a156f821a5b55554c2237.jpg"},{"id":71550443,"identity":"91e4e757-7f1c-4424-a1e5-833e16901168","added_by":"auto","created_at":"2024-12-16 15:39:51","extension":"docx","order_by":12,"title":"","display":"","copyAsset":false,"role":"supplement","size":18413,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryTable1.docx","url":"https://assets-eu.researchsquare.com/files/rs-5145248/v1/82625cd37072bb79b3fad648.docx"}],"financialInterests":"","formattedTitle":"Impact of Vitamin D3 Supplementation on Motor Functionality and the Immune Response in Parkinson's Disease Patients with Vitamin D Deficiency","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eParkinson\u0026apos;s disease, the second most prevalent neurodegenerative disorder, affects approximately 1\u0026ndash;2% of individuals aged 65 and older. Projections suggest that by 2040, the global prevalence of PD will exceed 12 million individuals worldwide. However, the exact etiology of PD remains unclear, with multiple factors implicated\u003csup\u003e1\u003c/sup\u003e. The intricate mechanism of neurodegeneration in PD has yet to be fully elucidated, but it is believed to encompass a complex interplay of genetic and environmental factors, oxidative stress, mitochondrial dysfunction, inflammation, immune modulation, and other factors.\u003c/p\u003e\n\u003cp\u003eVitamin D is a steroid hormone that performs many other functions\u0026nbsp;in addition to its well-known role in calcium homeostasis.\u0026nbsp;There is extensive evidence from in vitro and animal studies indicating that vitamin D plays crucial\u0026nbsp;roles\u0026nbsp;in cell proliferation and differentiation, neurotrophic regulation and neuroprotection, neurotransmission, immune regulation, and neural plasticity\u003csup\u003e2-6\u003c/sup\u003e. Furthermore, the expression of vitamin D receptor (VDR) and CYP27B1 is particularly abundant in the substantia nigra, a brain region rich in dopaminergic neurons\u003csup\u003e7\u003c/sup\u003e. As a fat-soluble hormone, vitamin D can cross the\u0026nbsp;blood‒brain\u0026nbsp;barrier, which serves as indirect evidence supporting its role in the central nervous system\u003csup\u003e8\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThere are four main subtypes of CD4+ T cells: Th1, Th2, Th17, and Treg cells. Among them,\u0026nbsp;Th17 cells are a proinflammatory subtype, whereas Treg cells are an anti-inflammatory subtype. The balance between Th17 and Treg cell levels plays a crucial role in maintaining immune homeostasis and inducing antigen-specific immune tolerance\u003csup\u003e9, 10\u003c/sup\u003e. Various autoimmune diseases are associated with the disruption of this balance\u003csup\u003e11\u003c/sup\u003e. Research over the past decade has shown that these immune alterations are not limited to the periphery but also extend to the central nervous system, where they promote\u0026nbsp;the progression of neurodegenerative diseases\u003csup\u003e12\u003c/sup\u003e. In cellular models, Th17 cells have been demonstrated to disrupt and cross the\u0026nbsp;blood‒brain\u0026nbsp;barrier and are associated with\u0026nbsp;increased\u0026nbsp;neuroinflammation in central nervous system pathologies\u003csup\u003e13, 14\u003c/sup\u003e. Within the central nervous system, regulatory T cells actively promote neural system recovery through the inhibition of astrocyte proliferation, with particular emphasis on their role in ischemic stroke and neuroinflammatory diseases\u003csup\u003e15\u003c/sup\u003e. Similar results have also been\u0026nbsp;reported\u0026nbsp;in studies involving PD patients, where Th17 cells were\u0026nbsp;found to induce\u0026nbsp;the\u0026nbsp;apoptosis of midbrain neurons\u003csup\u003e16\u003c/sup\u003e, whereas\u0026nbsp;Tregs exhibited\u0026nbsp;a protective effect on dopamine neurons\u003csup\u003e17\u003c/sup\u003e. Many studies have\u0026nbsp;reported\u0026nbsp;increased proportions of Th17 cells and decreased proportions and impaired\u0026nbsp;functions\u0026nbsp;of Treg cells in patients with Parkinson\u0026apos;s disease\u003csup\u003e18, 19\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eVitamin D can regulate Th17 and Treg cells primarily by inducing the expression of the \u003cem\u003eFoxp3\u003c/em\u003e gene in naive\u0026nbsp;CD4+\u0026nbsp;T\u0026nbsp;cells\u003csup\u003e20, 21\u003c/sup\u003e.\u0026nbsp;The\u0026nbsp;VDR binds to\u0026nbsp;\u003cem\u003eVDREs\u003c/em\u003e within the \u003cem\u003eFoxp3\u003c/em\u003e gene, enabling vitamin D to directly upregulate Foxp3 expression, thereby promoting the differentiation and expression of Treg cells. Conversely, Foxp3 inhibits the expression of ROR\u0026gamma;t in\u0026nbsp;CD4+T\u0026nbsp;cells\u003csup\u003e22\u003c/sup\u003e. ROR\u0026gamma;t is the primary transcription regulator of Th17 cells, promoting and maintaining their specificity as Th17 cells. Therefore, inhibiting ROR\u0026gamma;t expression leads to the suppression of Th17 differentiation and expression. In summary, vitamin D regulates the balance between Th17 and Treg\u0026nbsp;cells, playing a key role in maintaining immune homeostasis.\u003c/p\u003e\n\u003cp\u003eLow vitamin Dlevels in PD patients have been recognized for more than 15 years\u003csup\u003e23\u003c/sup\u003e, and numerous clinical studies have corroborated the imbalance of Th17 and Treg levels in these patients\u003csup\u003e18, 19, 24\u003c/sup\u003e. We hypothesized that it is possible to regulate the homeostatic balance between Th17 and Treg cells by improving low vitamin D levels in Parkinson\u0026apos;s disease patients, thereby improving clinical symptoms. This study aimed to detect the expression levels of vitamin D, Tregs, and Th17 cells in the peripheral blood of PD patients, explore the influence of Treg/Th17 imbalance on PD progression, and further investigate the impact of vitamin D intervention on this imbalance and clinical outcomes. We hope to gain a deeper understanding of the role of vitamin D deficiency and the Treg/Th17 imbalance in PD.\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cp\u003e2.1. Design of the Study\u003c/p\u003e\n\u003cp\u003eThis study was a randomized, double-blind, placebo-controlled clinical trial (NCT:\u0026nbsp;06539260) that administered a three-month supply of vitamin D\u003csub\u003e3\u003c/sub\u003e and a\u0026nbsp;placebo(vegetable oil)\u0026nbsp;to patients with PD who\u0026nbsp;had\u0026nbsp;low levels of vitamin D. This study obtained approval from the Ethics Committee of Suzhou Hospital of Anhui Medical University, with approval number A2023024. The procedures used in this study\u0026nbsp;adhered\u0026nbsp;to the principles of the Declaration of Helsinki. All patients signed informed consent to participate in the study and were informed of their right to withdraw at any point during the trial.\u003c/p\u003e\n\u003cp\u003e2.2. Participants\u003c/p\u003e\n\u003cp\u003eFrom January 2023 to March 2024, 51 PD patients were recruited. All the subjects were from the Parkinson\u0026apos;s outpatient clinic and inpatient ward of the Neurology Department at Suzhou Hospital of Anhui Medical University. Fifty partners or volunteers of PD patients, matched by sex\u0026nbsp;and age, were selected from Suzhou Hospital of Anhui Medical University to form a healthy control(HC) group.\u0026nbsp;The inclusion\u0026nbsp;criteria\u0026nbsp;for patients were as follows: (1)\u0026nbsp;met\u0026nbsp;the diagnostic criteria for PD established by the UK Parkinson\u0026apos;s Disease Society Brain Bank. (2) Age between 45 and 80 years. The exclusion\u0026nbsp;criteria\u0026nbsp;were as follows: (1) Having related vitamin D metabolic diseases (such as renal failure, severe liver damage, inherited 1\u0026alpha;-hydroxylase deficiency, etc.). (2) Having immune system diseases. (3) Having a history of disabling cerebrovascular disease. (4) Having first- or second-degree relatives with PD. (5) Having severe dementia, depression, or severe mental illness.\u003c/p\u003e\n\u003cp\u003e2.3. Randomization and Masking\u003c/p\u003e\n\u003cp\u003eFor the PD patients included in the study, analyses were conducted on their peripheral blood vitamin D levels. PD patients with low vitamin D levels were divided into two groups at a 1:1 ratio to receive vitamin D\u003csub\u003e3\u003c/sub\u003e and PL. Randomization was performed via an Excel random number generator. Both vitamin D\u003csub\u003e3\u003c/sub\u003e and the PL were placed in identical bottles labeled only with numbers and containing no additional information. Product allocation and randomization were carried out by an independent researcher. Both the personnel involved in data analysis and the patients remained blinded until the database was examined for analysis. Unblinding occurred when all the data were analyzed.\u003c/p\u003e\n\u003cp\u003e2.4. Outcome\u003c/p\u003e\n\u003cp\u003eThe primary endpoint of this study focused on the changes observed from baseline (T0) to the 3-month mark (T1). We evaluated PD patients via a comprehensive set of scales, including the Movement Disorder Society Unified Parkinson\u0026apos;s Disease Rating Scale (MDS-UPDRS), Berg Balance Scale (Breg), Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE), Self-rating Anxiety Scale (SAS), Self-rating Depression Scale (SDS), and Parkinson\u0026apos;s Disease Sleep Scale (PDSS).\u003c/p\u003e\n\u003cp\u003eDuring the initial T0 stage, we analyzed the peripheral blood levels of Th17, Treg, and Vit D in the PD patients and healthy volunteers. Among the PD patients with low vitamin D levels, we randomly assigned them into two groups: one to receive vitamin D\u003csub\u003e3\u003c/sub\u003e supplementation and the other to receive PL. At the T1 stage, we followed up with both groups of PD patients to reassess the indicators and monitor any changes (Fig. 1).\u003c/p\u003e\n\u003cp\u003e2.5. Measurement of vitamin D\u003c/p\u003e\n\u003cp\u003eThe serum 25(OH)D\u003csub\u003e3\u003c/sub\u003e concentration was measured via the Elecsys method on a Roche electrochemiluminescence fully automated immunoassay analyzer. According to the guidelines of the Endocrine Society, normal vitamin D levels are defined as \u0026ge;30 ng/mL, 20\u0026ndash;30 ng/mL is defined as insufficient, and vitamin deficiency is defined as \u0026le;20 ng/mL\u003csup\u003e25\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e2.6. Measurement of Th17 and Treg\u003c/p\u003e\n\u003cp\u003eVenous blood withdrawal was performed after a night of fasting, between 8:00 a.m. and 10:00 a.m., into heparin lithium anticoagulant tubes. The tubes were subsequently coded and stored at room temperature until processing, which occurred 12 hours after collection, to ensure homogeneous treatment of all the samples.\u003c/p\u003e\n\u003cp\u003eTh17 cells by flow cytometry\u003c/p\u003e\n\u003cp\u003eThe blood samples were diluted 1:1 with high-sugar medium (RPMI 1640) and mixed well. Leukocyte activation cocktail (3 \u0026mu;L/1 ml, BD Pharmingen) was added to the solution of diluted blood. The mixture was incubated for 4\u0026ndash;6 hours in 5% CO2 at 37\u0026deg;C. Two milliliters of hemolysin (BD Pharmingen) was added, and the mixture was vortexed and incubated for 10 minutes at room temperature in the dark. The samples were subsequently centrifuged, the supernatants were removed, the cells were washed in PBS, and the resulting mixture was resuspended in PBS. One hundred microliters of PBS buffer was added, the cells were resuspended, FITC-conjugated mouse anti-human CD3 (BD Pharmingen) and CD8-conjugated PERCP-CY5.5 (BioLegend) were added for surface antibody staining, and the mixture was incubated for 15 minutes in the dark. After the cells were fixed and permeabilized (Fixation/Permeabilization Kit, BD Pharmingen), an intracellular PE-conjugated mouse anti-human IL-17A antibody (BD Pharmingen) was added, and the mixture was incubated at room temperature in the dark for 30 minutes.\u003c/p\u003e\n\u003cp\u003eAssessment of Tregs by flow cytometry\u003c/p\u003e\n\u003cp\u003eCD4-FITC and CD25-PE antibodies (BD Pharmingen) were added to the flow tube. Then, 50 \u0026micro;l of peripheral blood was added to the tube, which was vortexed and mixed well. The mixture was incubated in the dark for 15 minutes. Then, 500 \u0026mu;L of hemolysin (BD Pharmingen) was added, the mixture was shaken, and the mixture was incubated in the dark for 15 minutes. Three milliliters of PBS was added, the mixture was centrifuged for 5 minutes, and the supernatant was removed. The cells were fixed and permeabilized (FOXP3/TRN FACTOR STAIN BUFFER, eBioscience). The intranuclear staining antibody foxp3 (ANTI-HU FOXP3 PCH101 APC, eBioscience) was added to each tube, and the mixture was incubated at 4\u0026deg;C for 2 hours in the dark. Three milliliters of PBS was added again, the mixture was centrifuged for 5 minutes, and the supernatant was removed.\u003c/p\u003e\n\u003cp\u003eAcquisition was then performed on a BD FACSCanto II flow cytometer. CD3\u003csup\u003e+\u003c/sup\u003eCD8\u003csup\u003e-\u003c/sup\u003eIL-17\u003csup\u003e+\u003c/sup\u003e cells were identified as Th17 cells, whereas CD4+CD25+Foxp3+ cells were identified as Treg cells. Lymphocytes are recognized on the basis of their classical forward scatter (FSC) and side scatter (SSC) signals. The data were analyzed via FlowJo software (version 10.8.1).\u003c/p\u003e\n\u003cp\u003e2.7 Statistical analysis\u003c/p\u003e\n\u003cp\u003eIn this study, SPSS 26.0 and GraphPad Prism v.9 software were used for data analysis. Descriptive statistical methods were employed to analyze all the data gathered during the research process. For intergroup comparisons, an independent sample t test was conducted, \u0026nbsp;a paired sample t test was used for comparisons before and after the follow-up. Pearson correlation analysis was applied to data adhering to a normal distribution, whereas Spearman correlation analysis was employed for data deviating from a normal distribution. A P value of less than 0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"3. Results","content":"\u003cp\u003e3.1 Demographic characteristics\u003c/p\u003e\n\u003cp\u003eThis study included a total of 51 PD patients and 50 healthy individuals, all of whom were of Chinese ethnicity and fulfilled the established inclusion criteria. The PD group included 29 males and 22 females, with a mean age of 69.6\u0026plusmn;11.0 years. The HCs included 26 males and 24 females, with a mean age of 65.3\u0026plusmn;10.9 years. No statistically significant differences were observed in sex composition or age between the two groups (P\u0026gt;0.05). The PD group presented a mean H-Y grade of 2.32\u0026plusmn;0.90, an average UPDRS score of 45.76\u0026plusmn;23.06, a mean UPDRS-III score of 26.35\u0026plusmn;13.56, an average Breg score of 43.45\u0026plusmn;12.18, a mean MMSE score of 21.46\u0026plusmn;7.07, an average MoCA score of 18.32\u0026plusmn;7.51, a mean SDS self-rating depression scale score of 45.85\u0026plusmn;13.47, a mean SAS self-rating anxiety scale score of 44.20\u0026plusmn;12.50, and an average PDSS score of 100.82\u0026plusmn;23.48, as indicated in Table 1. The PD patients enrolled were categorized into a normal vitamin D level group (Vit D\u0026ge;30 ng/ml), consisting of 21 patients, and a low vitamin D level group (Vit D\u0026lt;30 ng/ml), comprising 30 patients, on the basis of their baseline vitamin D levels. The low vitamin D group was subsequently randomly divided into two subgroups, a VitD\u003csub\u003e3\u003c/sub\u003e group and a PL group, each comprising 15 patients. Importantly, no complications arose during or following vitamin D supplementation for any of the subjects with vitamin D deficiency.\u003c/p\u003e\n\u003cp\u003e3.2 Comparison of 25(OH) D\u003csub\u003e3\u003c/sub\u003e, Th17, and Treg cells in the PD Group and HC Group\u003c/p\u003e\n\u003cp\u003eTo investigate changes in CD4+ T-cell subsets in the peripheral blood, we first evaluated the percentages of CD4+ T-cell subsets (Th17 and Treg) in 51 PD patients and 50 healthy controls. Compared with the HCs, the PD cohort presented significant differences in Th17 levels (3.78\u0026plusmn;1.33 vs 2.30\u0026plusmn;0.79, t=6.799, p\u0026lt;0.001; Fig. 2a) and Treg levels (4.16\u0026plusmn;1.29 vs 4.64\u0026plusmn;0.97, t=-2.142, p=0.035; Fig. 2b). Additionally, there was a significant difference in the serum 25(OH) D\u003csub\u003e3\u003c/sub\u003e level between the PD cohort and the HC cohort (28.98\u0026plusmn;8.10 vs 35.80\u0026plusmn;6.96, t=-4.531; p\u0026lt;0.001; Fig. 2c).\u003c/p\u003e\n\u003cp\u003e3.3 Vitamin D, Th17, and Treg correlations in human blood\u003c/p\u003e\n\u003cp\u003eWe conducted a correlation analysis on a total of 101 data sets from the PD group and the HC group. The analysis revealed a positive correlation between vitamin D levels in peripheral blood and the percentage of Tregs (r=0.526, p\u0026lt;0.001; Fig. 3a) and a\u0026nbsp;negative correlation with the percentage of Th17 cells (r=-0.635, p\u0026lt;0.001; Fig. 3b).\u003c/p\u003e\n\u003cp\u003e3.4 Correlations between PD-related scores and Th17, Treg, and vitamin D levels\u003c/p\u003e\n\u003cp\u003eThe proportion of Th17 cells was positively correlated with both the UPDRS and the UPDRS-III score (r=0.412, p=0.003 and r=0.432, p=0.002, respectively) and negatively correlated with the Breg balance scale (r=-0.302, P=0.031). The proportion of Treg cells was negatively correlated with the UPDRS and UPDRS-III scores (r=-0.504, p\u0026lt;0.001 and r=-0.540, p\u0026lt;0.001, respectively) and positively correlated with the Breg balance scale (r=0.382, p=0.006). A correlation analysis between vitamin D levels and UPDRS and UPDRS-III scores among PD patients revealed a negative correlation (r=-0.494, p\u0026lt;0.001 and r=-0.549, p\u0026lt;0.001, respectively) and a positive correlation with Breg (r=0.419, p=0.002), as indicated in\u0026nbsp;Table 2. These results indicate that the severity of motor impairment in Parkinson\u0026apos;s disease patients is correlated with the levels of Th17, Treg, and vitamin D. The MMSE score of PD patients is positively correlated with Th17 levels but negatively correlated with vitamin D levels. The MoCA score is positively correlated with Th17 levels and negatively correlated with Treg and vitamin D levels. On the other hand, no correlation was observed between the SDS and SAS scores of PD patients and the levels of Th17, Treg, and vitamin D. Additionally, the PDSS score positively correlated with Treg but was not correlated with Th17 and vitamin D, as indicated in Table 2.\u0026nbsp;Therefore, the intellectual status of PD patients seems to be associated with the levels of Th17, Treg, and vitamin D.\u003c/p\u003e\n\u003cp\u003e3.5 Follow-up results\u003c/p\u003e\n\u003cp\u003eThere were 30 patients with low vitamin D levels in the PD group; these patients were randomly divided into the VitD\u003csub\u003e3\u003c/sub\u003e subgroup 15 patients and the PL subgroup 15 patients. The basic characteristics of the two subgroups are shown in Table 3.\u0026nbsp;After a three-month follow-up with medication administration, the VitD\u003csub\u003e3\u003c/sub\u003e subgroup exhibited a notable reduction in Th17 levels during vitamin D supplementation, with levels decreasing from 4.62\u0026plusmn;1.09 to 3.25\u0026plusmn;1.14 (P=0.003, Fig. 4a). In contrast, Th17 levels did not significantly change in the PL subgroup before or after the three-month period (p\u0026gt;0.05, Fig. 4a). The Treg levels in the VitD\u003csub\u003e3\u003c/sub\u003e subgroup increased (3.25\u0026plusmn;0.90 vs 4.52\u0026plusmn;0.95, P=0.003; Fig. 4b), whereas there was no significant difference in the Treg levels before and after treatment in the PL subgroup (p\u0026gt;0.05; Fig. 4b). Additionally, we evaluated the VitD\u003csub\u003e3\u003c/sub\u003e subgroup of PD patients via the UPDRS, UPDRS-III, and Breg scales to assess three motor indicators before and after treatment. We found that the VitD\u003csub\u003e3\u003c/sub\u003e subgroup showed significant improvement in motor indicators after three months of vitamin D supplementation, with UPDRS scores improving from 57.00\u0026plusmn;20.86 to 52.27\u0026plusmn;21.38 (P=0.003, Fig. 4c) and UPDRS-III scores improving from 32.40\u0026plusmn;11.70 to 28.13\u0026plusmn;12.44 (P\u0026lt;0.001, Fig. 4d). However, there was no difference in the Breg scale score before and after treatment (39.73\u0026plusmn;13.36 vs 40.60\u0026plusmn;12.91, P=0.400; Fig. 4e). Furthermore, there were no differences in motor indicators before and after three months in the PL subgroup, as indicated in Supplementary Table 1.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eOur findings indicate that the average vitamin D level in PD patients is lower than that in healthy controls, which is consistent with the results of several previous clinical studies\u003csup\u003e23\u003c/sup\u003e. Additionally, PD patients have a higher Th17/Treg ratio than healthy individuals do. Upon comprehensive analysis, we observed a significant correlation between the levels of 25(OH) D\u003csub\u003e3\u003c/sub\u003e in human peripheral blood and Th17 and Treg cells. This finding is in line with our hypothesis that 25-hydroxyvitamin D\u003csub\u003e3\u003c/sub\u003e interacts with the vitamin D receptor on the surface of CD4+ T cells, participating in their differentiation and expression and promoting the expression and differentiation of Treg cells while inhibiting the differentiation and expression of Th17 cells. For the analysis of motor function in PD patients, we used the UPDRS-III and Breg balance scales for assessment. There was a significant correlation between the UPDRS-III and Breg scores of PD patients and the levels of Th17, Treg, and 25(OH) D\u003csub\u003e3\u003c/sub\u003e. Furthermore, our analysis of nonmotor scales revealed a significant correlation between the cognitive scales (MoCA, MMS) of PD patients and the levels of Th17, Treg, and\u0026nbsp;25(OH) D\u003csub\u003e3\u003c/sub\u003e.\u003c/p\u003e\n\u003cp\u003eA three-month follow-up was conducted on both the VitD\u003csub\u003e3\u003c/sub\u003e group and the PL group. The results revealed that the levels of 25(OH) D\u003csub\u003e3\u003c/sub\u003e in the peripheral blood of the VitD\u003csub\u003e3\u003c/sub\u003e group were increased, and the levels of Th17 and Treg cells tended toward those of the normal population. Furthermore, compared with their preintervention levels and those of the PL group at the three-month follow-up, the motor function of the VitD\u003csub\u003e3\u003c/sub\u003e group had improved.\u003c/p\u003e\n\u003cp\u003eThe role of vitamin D in PD has been extensively studied. In some studies, vitamin D deficiency has been observed among PD patients\u003csup\u003e26\u003c/sup\u003e. Research has revealed a correlation between the severity of motor impairments in PD patients and their peripheral blood vitamin D levels\u003csup\u003e27, 28\u003c/sup\u003e, and our study yielded similar results. In 2010, a longitudinal study based on a health survey of the Finnish population revealed that individuals with low vitamin D levels had a 65% greater probability of developing Parkinson\u0026apos;s disease than did those with high vitamin D levels\u003csup\u003e29\u003c/sup\u003e. These findings indirectly support the role of vitamin D in the onset and progression of PD. Vitamin D can achieve neuroprotective effects by inhibiting the early aggregation of \u0026alpha;-synuclein (\u0026alpha;-Syn), especially in\u0026nbsp;neurodegenerative diseases\u003csup\u003e30\u003c/sup\u003e.\u0026nbsp;A\u0026nbsp;study on the treatment of Parkinson\u0026apos;s disease in animal models with vitamin D\u0026nbsp;revealed\u0026nbsp;that vitamin D\u0026nbsp;has\u0026nbsp;neuroprotective effects by attenuating\u0026nbsp;proinflammatory\u0026nbsp;processes and\u0026nbsp;upregulating\u0026nbsp;anti-inflammatory processes in animal models of PD\u003csup\u003e31\u003c/sup\u003e. Vitamin D mediates the secretion and expression of inflammatory factors, which may be the pathway through which it influences the onset and progression of PD. In 1988, McGeer et al. discovered HLA-DR-reactive microglia in the postmortem substantia nigra tissue of Parkinson\u0026apos;s disease patients\u003csup\u003e32\u003c/sup\u003e, which was one of the earliest\u0026nbsp;lines of evidence\u0026nbsp;linking neuroinflammation to the pathogenesis of Parkinson\u0026apos;s disease.\u0026nbsp;Microglia, monocytes, and infiltrating T\u0026nbsp;cells,\u0026nbsp;which migrate\u0026nbsp;around \u0026alpha;-syn\u0026nbsp;in PD patients\u0026apos; substantia nigra, suggest both\u0026nbsp;the role of\u0026nbsp;innate and adaptive\u0026nbsp;immunity\u0026nbsp;in PD\u003csup\u003e33\u003c/sup\u003e.\u0026nbsp;A recent study\u0026nbsp;revealed\u0026nbsp;that \u0026alpha;-Syn levels in PD patients are associated with the severity of motor symptoms and\u0026nbsp;an\u0026nbsp;imbalance of Th17/Treg\u0026nbsp;cells\u003csup\u003e18\u003c/sup\u003e. Furthermore,\u0026nbsp;a previous\u0026nbsp;study suggested that \u0026alpha;-Syn impairs the stability of Tregs and promotes the differentiation of Th17 cells in PD\u003csup\u003e18\u003c/sup\u003e.\u0026nbsp;Contact\u0026nbsp;between Th17 cells and midbrain neurons can directly lead to the apoptosis of dopaminergic neurons\u003csup\u003e16, 34\u003c/sup\u003e, and IL-17 has a destructive effect on the\u0026nbsp;blood‒brain\u0026nbsp;barrier\u003csup\u003e35\u003c/sup\u003e. Conversely, Treg cells can significantly protect the survival of midbrain dopaminergic neurons\u003csup\u003e17\u003c/sup\u003e, which also indirectly indicates that\u0026nbsp;an\u0026nbsp;imbalance of Th17/Treg\u0026nbsp;cells\u0026nbsp;in PD patients may exacerbate clinical symptoms. Treg cells play a significant role in reducing\u0026nbsp;the\u0026nbsp;beta-amyloid protein load in Alzheimer\u0026apos;s patients, restoring brain homeostasis, and improving learning and memory\u003csup\u003e36\u003c/sup\u003e. Similarly, our study\u0026nbsp;revealed\u0026nbsp;a clear correlation between Treg levels and cognitive function in PD patients, suggesting that improving Treg levels may be a potential therapeutic approach for PD patients with cognitive issues.\u003c/p\u003e\n\u003cp\u003e\u0026alpha;-Syn\u0026nbsp;promotes the differentiation of Th17 cells and activates intracellular inflammatory pathways through autoimmune effects\u003csup\u003e18\u003c/sup\u003e, and low vitamin D levels may exacerbate this situation\u003csup\u003e30\u003c/sup\u003e. Either through the direct immunomodulatory effects of vitamin D or through the immune pathways mediated by \u0026alpha;-syn, low vitamin D levels lead to an imbalance between Th17 cells and Treg cells\u003csup\u003e37\u003c/sup\u003e, causing damage to substantia nigra neurons\u003csup\u003e16, 17\u003c/sup\u003e. We believe that this pathway represents a promising direction for future research on the pathogenesis of Parkinson\u0026apos;s disease and may lead to the discovery of new treatments.\u003c/p\u003e\n\u003cp\u003eThis study has several limitations. First, the relatively small sample size restricts the generalizability of the results to a limited scope. Second, the observed vitamin D deficiency in PD patients could stem from reduced outdoor activity and inadequate sunlight exposure due to motor disorders, subsequently hindering vitamin D synthesis. However, we have not undertaken any supplementary analysis specifically addressing this underlying mechanism.\u0026nbsp;Third, while our study\u0026nbsp;revealed\u0026nbsp;improvements in Th17 and Treg levels, as well as motor function, among PD patients receiving vitamin D supplementation, we remain unable to definitively attribute the motor improvement solely to the restoration of neural function resulting from the improved Th17/Treg balance.\u003c/p\u003e\n\u003cp\u003eIn the future, further experiments are needed to explore the deeper mechanisms by which vitamin D affects the balance between Th17 and Treg cells and to verify the neuroprotective effect of vitamin D on PD patients through improvements in their autoimmune status.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eIn the present study, we observed that, compared with healthy individuals, PD patients presented elevated levels of Th17 and decreased levels of Treg and vitamin D.\u0026nbsp;The more pronounced these numerical deviations are, the more severe the motor dysfunction symptoms become in PD patients. Supplementing PD patients with low vitamin D levels could regulate their Th17 and Treg levels, reduce autoimmune inflammation, and improve motor function.\u0026nbsp;In summary, we believe that restoring vitamin D levels to normal is necessary for PD patients with vitamin D deficiency, as it can\u0026nbsp;increase\u0026nbsp;their self-care\u0026nbsp;ability\u0026nbsp;and help alleviate or slow the progression of disability. Our data\u0026nbsp;support\u0026nbsp;the notion that vitamin D plays a crucial role in mediating the balance between Th17 and Treg\u0026nbsp;cells, which is closely linked to the pathogenesis of Parkinson\u0026apos;s disease.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAcknowledgments\u003c/p\u003e\n\u003cp\u003eThe authors have no acknowledgments to report.\u003c/p\u003e\n\u003cp\u003eFunding Statement\u003c/p\u003e\n\u003cp\u003eThis research was supported by the Scientific Research Project of the Health Commission of Anhui Province (AHWJ2022b106) and the Anhui Provincial Key Research and Development Plan (202204295107020063).\u003c/p\u003e\n\u003cp\u003eConflict of interest\u003c/p\u003e\n\u003cp\u003eAll the authors declare that this research was conducted without any commercial or financial relationships that could be construed as potential conflicts of interest.\u003c/p\u003e\n\u003cp\u003eData availability statement\u003c/p\u003e\n\u003cp\u003eThe data supporting the findings of this study are available within the article and its supplementary material.\u003c/p\u003e\n\u003cp\u003eEthical approval\u003c/p\u003e\n\u003cp\u003eThis study was conducted in accordance with ethical principles that originated in the Declaration of Helsinki and that are consistent with good clinical practice. Institutional review boards or independent ethics committees provided written approval for the study protocol and all amendments. All patients provided written informed consent.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eClinicalTrials.gov: NCT:06539260. Registered 05 August 2024 - Retrospectively registered, https://clinicaltrials.gov/study/NCT06539260.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAuthor contributions\u003c/p\u003e\n\u003cp\u003eDanfeng Li\u0026nbsp;and Xibo Ma\u0026nbsp;wrote this manuscript. Yan Chen, Xiaowei Zhu, and Xibo Ma participated in data collection. Wentao Zhang and Danfeng Li analyzed the data. Shihua Liu and Ping Zhong guided and revised the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eDorsey ER, Sherer T, Okun MS, et al. The Emerging Evidence of the Parkinson Pandemic. \u003cem\u003eJ Parkinsons Dis\u003c/em\u003e 2018; 8: S3-S8. 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DOI: 10.1210/jc.2011-0385.\u003c/li\u003e\n\u003cli\u003eZhou Z, Zhou R, Zhang Z, et al. The Association Between Vitamin D Status, Vitamin D Supplementation, Sunlight Exposure, and Parkinson's Disease: A Systematic Review and Meta-Analysis. \u003cem\u003eMed Sci Monit\u003c/em\u003e 2019; 25: 666-674. 20190123. DOI: 10.12659/MSM.912840.\u003c/li\u003e\n\u003cli\u003eSuzuki M, Yoshioka M, Hashimoto M, et al. 25-hydroxyvitamin D, vitamin D receptor gene polymorphisms, and severity of Parkinson's disease. \u003cem\u003eMov Disord\u003c/em\u003e 2012; 27: 264-271. 20111231. DOI: 10.1002/mds.24016.\u003c/li\u003e\n\u003cli\u003eDing. H, Dhima. K, Lockhart. KC, et al. Unrecognized vitamin D3 deficiency is common in Parkinson disease. \u003cem\u003eNeurology\u003c/em\u003e 2013: 1531\u0026ndash;1537. DOI: 10.1212/WNL.0b013e3182a95818.\u003c/li\u003e\n\u003cli\u003eKnekt P, Kilkkinen A, Rissanen H, et al. Serum vitamin D and the risk of Parkinson disease. \u003cem\u003eArch Neurol\u003c/em\u003e 2010; 67: 808-811. DOI: 10.1001/archneurol.2010.120.\u003c/li\u003e\n\u003cli\u003eZhang Y, Ji W, Zhang S, et al. Vitamin D Inhibits the Early Aggregation of alpha-Synuclein and Modulates Exocytosis Revealed by Electrochemical Measurements. \u003cem\u003eAngew Chem Int Ed Engl\u003c/em\u003e 2022; 61: e202111853. 20211122. DOI: 10.1002/anie.202111853.\u003c/li\u003e\n\u003cli\u003eCalvello R, Cianciulli A, Nicolardi G, et al. Vitamin D Treatment Attenuates Neuroinflammation and Dopaminergic Neurodegeneration in an Animal Model of Parkinson's Disease, Shifting M1 to M2 Microglia Responses. \u003cem\u003eJ Neuroimmune Pharmacol\u003c/em\u003e 2017; 12: 327-339. 20161216. DOI: 10.1007/s11481-016-9720-7.\u003c/li\u003e\n\u003cli\u003eMcGeer PL, Itagaki S and BE. B. Reactive microglia are positive for HLA-DR in the substantia nigra of Parkinson\u0026rsquo;s and Alzheimer\u0026rsquo;s disease brains. \u003cem\u003eNeurology\u003c/em\u003e 1988; 38: 1285-1291. DOI: 10.1212/wnl.38.8.1285.\u003c/li\u003e\n\u003cli\u003eTansey MG, Wallings RL, Houser MC, et al. Inflammation and immune dysfunction in Parkinson disease. \u003cem\u003eNat Rev Immunol\u003c/em\u003e 2022; 22: 657-673. 20220304. DOI: 10.1038/s41577-022-00684-6.\u003c/li\u003e\n\u003cli\u003eLindestam Arlehamn CS, Dhanwani R, Pham J, et al. alpha-Synuclein-specific T cell reactivity is associated with preclinical and early Parkinson's disease. \u003cem\u003eNat Commun\u003c/em\u003e 2020; 11: 1875. 20200420. DOI: 10.1038/s41467-020-15626-w.\u003c/li\u003e\n\u003cli\u003eLiu Z, Qiu AW, Huang Y, et al. IL-17A exacerbates neuroinflammation and neurodegeneration by activating microglia in rodent models of Parkinson's disease. \u003cem\u003eBrain Behav Immun\u003c/em\u003e 2019; 81: 630-645. 20190724. DOI: 10.1016/j.bbi.2019.07.026.\u003c/li\u003e\n\u003cli\u003eYeapuri P, Machhi J, Lu Y, et al. Amyloid-beta specific regulatory T cells attenuate Alzheimer's disease pathobiology in APP/PS1 mice. \u003cem\u003eMol Neurodegener\u003c/em\u003e 2023; 18: 97. 20231218. DOI: 10.1186/s13024-023-00692-7.\u003c/li\u003e\n\u003cli\u003eAo T, Kikuta J and Ishii M. The Effects of Vitamin D on Immune System and Inflammatory Diseases. \u003cem\u003eBiomolecules\u003c/em\u003e 2021; 11 20211103. DOI: 10.3390/biom11111624.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1 Characteristics of PD patients and healthy controls.\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"99%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 31.3131%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.2929%;\"\u003e\n \u003cp\u003ePD Group\u003c/p\u003e\n \u003cp\u003e(n=51)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 39.3939%;\"\u003e\n \u003cp\u003eHC Group\u003c/p\u003e\n \u003cp\u003e(n=50)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003eM: 29\u003c/p\u003e\n \u003cp\u003eW: 22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003eM: 26\u003c/p\u003e\n \u003cp\u003eW: 24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e69.6\u0026plusmn;11.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e65.3\u0026plusmn;10.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eH-Y\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e2.32\u0026plusmn;0.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eUPDRS\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e45.76\u0026plusmn;23.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eUPDRS-III\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e26.35\u0026plusmn;13.56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eBreg\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e43.45\u0026plusmn;12.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eMMSE\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e21.46\u0026plusmn;7.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eMoCA\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e18.32\u0026plusmn;7.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eSDS\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e45.85\u0026plusmn;13.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003eSAS\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e44.20\u0026plusmn;12.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3131%;\"\u003e\n \u003cp\u003ePDSS\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29.2929%;\"\u003e\n \u003cp\u003e100.82\u0026plusmn;23.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.3939%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable 2 Correlations between PD-related scores and Th17, Treg, and vitamin D levels.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTh17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTreg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eVitamin D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003er\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003er\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003er\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eUPDRS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.412\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.504\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.494\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eUPDRS-III\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.432\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.540\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.549\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eBreg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.302\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.031\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.382\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.006\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.419\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eMMSE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.352\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.161\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.263\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eMoCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.377\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.308\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.029\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.374\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSDS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.933\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.148\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.299\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.450\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSAS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.111\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.440\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.210\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.138\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.224\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.114\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003ePDSS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.050\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.727\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.347\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.013\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-0.137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.338\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable 3 Characteristics of the VitD group and PL group.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 24px;\"\u003e\n \u003cp\u003eVitD\u003c/p\u003e\n \u003cp\u003e(n=15)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 27px;\"\u003e\n \u003cp\u003ePL\u003c/p\u003e\n \u003cp\u003e(n=15)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003ep\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003eM: 9\u003c/p\u003e\n \u003cp\u003eW: 6\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003eM: 9\u003c/p\u003e\n \u003cp\u003eW: 6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e73.4\u0026plusmn;8.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e73.1\u0026plusmn;10.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eH-Y\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e2.63\u0026plusmn;0.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e2.67\u0026plusmn;0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eTh17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e4.62\u0026plusmn;1.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e4.41\u0026plusmn;1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eTreg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e3.25\u0026plusmn;0.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e3.61\u0026plusmn;0.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eVitamin D\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e22.98\u0026plusmn;3.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e23.84\u0026plusmn;3.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eUPDRS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e57.0\u0026plusmn;20.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e54.4\u0026plusmn;25.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eUPDRS-III\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e32.4\u0026plusmn;11.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e-34.0\u0026plusmn;14.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eBreg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24px;\"\u003e\n \u003cp\u003e39.7\u0026plusmn;13.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e37.3\u0026plusmn;13.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Parkinson's disease, Th17, Treg, vitamin D","lastPublishedDoi":"10.21203/rs.3.rs-5145248/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5145248/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The purpose of this study was to investigate the levels of vitamin D, regulatory T (Treg) cells, and T helper 17 (Th17) cells in the peripheral blood of patients with Parkinson's disease(PD), explore their potential relationships with PD, and further examine the effects of vitamin D intervention on these biomarkers and clinical outcomes. PD patients with low vitamin D3 levels were randomly assigned to two groups: those supplemented with vitamin D3 (VitD3, n=15) and those treated with vegetable oil (PL, n=15). Treatment was administered continuously for 3 months. Vitamin D3, Th17 and Treg levels and related clinical scales were continuously monitored and evaluated. The results revealed that the level of 25(OH) D3 in the peripheral blood increased in the VitD3 group, and the Th17 and Treg levels tended toward those of the normal population. Compared with those of the preintervention groups and the PL group after the 3-month follow-up, motor function improved in the VitD3 group. Additionally, vitamin D was negatively correlated with Th17 cells and positively correlated with Treg cells. In PD patients, motor function was positively correlated with vitamin D and Treg levels but negatively correlated with Th17 levels. We believe that it is necessary for PD patients with low vitamin D levels to supplement vitamin D to normal levels.","manuscriptTitle":"Impact of Vitamin D3 Supplementation on Motor Functionality and the Immune Response in Parkinson's Disease Patients with Vitamin D Deficiency","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-16 15:39:46","doi":"10.21203/rs.3.rs-5145248/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":"6d4e8abd-fb01-4d12-9c79-63585ad3248b","owner":[],"postedDate":"December 16th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-25T18:50:43+00:00","versionOfRecord":[],"versionCreatedAt":"2024-12-16 15:39:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5145248","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5145248","identity":"rs-5145248","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}