Association between Cytokines, 25-Hydroxyvitamin D, and Physical Activity: Evidence from a  cross sectional Brazilian Population

Association between Cytokines, 25-Hydroxyvitamin D, and Physical Activity: Evidence from a cross sectional Brazilian Population | 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 Association between Cytokines, 25-Hydroxyvitamin D, and Physical Activity: Evidence from a cross sectional Brazilian Population Giana Zarbato Longo, Danielle Cristina Guimarães da Silva, Mariana Papinni Gabiatti, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6214919/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Vitamin D is of vital importance for bone health and also regulates inflammatory cytokines, which contribute to immune signaling and defenses. It is possible that physical activity could influence serum 25(OH)D levels, and it has been suggested that exercise can increase serum 25(OH)D concentration. The aim of this study was to investigate the association between inflammatory biomarkers and 25(OH)D concentration in a sample of Brazilian adults, taking into account the influence of physical activity on this association. Methods: This is a cross-sectional population-based study, conducted with 712 adults aged 20 to 59 years living in the urban area of Viçosa, Minas Gerais, Brazil. Sociodemographic, anthropometric, behavioral and biochemical variables were collected. Multiple linear regression models (crude and adjusted) were used to determine associations between 25(OH)D concentration and inflammatory biomarkers, with statistical significance analyzed at 5%. Results: In the multiple linear regression models, it can be observed that for each unit increase in the concentration of 25(OH)D, there was an average decrease in the inflammatory biomarkers. Physical activity level was considered an effect modifier of the association between serum 25(OH)D levels and some cytokines, and participants whose PAL was greater than 150 minutes tended to have lower levels of IL-6, IL-10, TNF-α and IL-12p70 when plasma 25(OH)D concentration was increased. Conclusions: Higher 25(OH)D concentrations were associated with lower levels of inflammatory biomarkers in Brazilian adults. Furthermore, physical activity served as a significant effect modifier of this association, enhancing the reduction of pro-inflammatory cytokines. These results underscore the importance of maintaining adequate 25(OH)D levels in conjunction with an active lifestyle to modulate inflammation and promote overall health. 25-hydroxyvitamin D cytokines inflammation physical activity nutritional epidemiology. Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Vitamin D (25(OH)D) deficiency is a global problem and is prevalent in tropical countries[ 1 ]. In Brazil, a meta-analysis aimed at studying the prevalence of deficiency and insufficiency of this vitamin in the Brazilian population in different age groups found values of 28.16% (95% CI: 23.90, 32.40) and 45.26% (95% CI: 35.82, 54.71)[ 2 ], respectively, confirming that hypovitaminosis D is a public health problem. Research conducted in recent years has shown the association between hypovitaminosis D and several diseases, including cancer[ 3 ], cardiometabolic disease[ 4 ], and inflammatory disorders[ 5 ], and has highlighted the association of this vitamin with some inflammatory cytokines and immune signaling, attracting increasing attention from the scientific community. 25(OH)D can reduce systemic inflammatory mediators and promote the release of anti-inflammatory cytokines from the immune system. A possible explanation for this relationship may be the anti-inflammatory properties of 25(OH)D, which affects the production and function of adipokines, thereby modulating the inflammatory response in adipose tissue[ 4 ]. In the NHANES study, in a probabilistic sample of US adults, individuals with high serum concentrations of 25(OH)D were found to have a better profile of inflammatory biomarkers[ 6 ]. Few studies have investigated the relationship between the effect of 25(OH)D in combination with physical activity throughout the seasons[ 7 – 9 ]. The interaction between physical activity and serum 25(OH)D levels has been explored by several studies, suggesting physiological mechanisms that explain this relationship and the potential effect on inflammation modulation. One of the main hypotheses is that physical activity stimulates lipolysis in adipose tissue, releasing the vitamin D stored there and consequently increasing serum 25(OH)D concentrations[ 10 ]. In addition, regular physical activity is associated with an improved metabolic profile, reduced body adiposity, and maintenance of a healthy weight, factors that may also positively influence vitamin D status[ 11 ]. From an immunological point of view, regular physical activity stimulates the production of anti-inflammatory myokines (e.g. IL-10) and reduces the concentration of pro-inflammatory cytokines (e.g. TNF-α), thus contributing to a more balanced immune environment[ 12 , 13 ]. Therefore, when combined, adequate levels of 25(OH)D and physical activity can potentiate the reduction of systemic inflammatory markers. Considering the possible modulating effect of 25(OH)D) on inflammation and its influence on the reduction of inflammatory markers associated with a sufficient level of physical activity, our hypothesis is that higher plasma levels of 25(OH)D are associated with lower plasma concentrations of cytokines in physically active individuals. Therefore, the aim of this study was to investigate the association between inflammatory biomarkers and 25(OH)D concentration in a sample of Brazilian adults, taking into account the influence of physical activity on this association. Materials and methods Study design and participants This is a population-based cross-sectional study aimed at assessing the health status of the adult population of Viçosa/MG, Brazil (2012–2014). The sample was composed of non-institutionalized individuals of both sexes, between 20 and 59 years of age, who were residents of the urban area of the city. The sampling process was probabilistic without repositioning in two stages (census sectors and domicile). In the end, 1229 home interviews were conducted; 331 of the interviewees did not complete the laboratory examination and 186 were excluded from the sample due to loss of the biological sample. The final sample size was 712 (Fig. 1 ). There were no statistical differences in sociodemographic characteristics between the original sample and the sample used in this study (data not shown). This study was conducted in accordance with the Declaration of Helsinki. The research project was approved by the Research Ethics Committee of the Federal University of Viçosa (Official Letter 02/2013). All participants signed the informed consent form. Additional methodological details can be found in Segheto et al. (2015)[ 14 ]. Sociodemographic, behavioral and seasonality variables The sociodemographic variables were: age, expressed as completed years and age group (20–39; 40–59 years); sex (male and female); years of schooling completed. The behavioral variables were: cigarette smoking (current smoker, ex-smoker and never-smoker) regardless of frequency and intensity of tobacco use[ 15 ]; and physical activity level (PAL). PAL was assessed using the long version (version 6) of the International Physical Activity Questionnaire (IPAQ) as described by Matsudo et al. (2001) [ 16 , 17 ]. For this study, only time spent in leisure-time physical activity during the week prior to administration of the questionnaire was considered. Participants were then categorized as irregularly active (IA) (< 150 min/week) or physically active (PA) (≥ 150 min/week)[ 16 ] (World Health Organization, 2020). The seasonality variable, which indicates the time of year when blood was collected to assess vitamin D status, was dichotomized into winter and summer/spring/autumn. Anthropometric parameters Body mass index (BMI) was calculated as body mass (kg) divided by height (m) squared, expressed in kilograms per meter squared (kg/m 2 ). Body mass was measured with a digital scale (Tanita, Ironman TM model, BC-554, Tanita Corporation), and height was measured using a metal stadiometer (Welmy, Santa Bárbara D'Oeste, SP, Brazil). During the anthropometric evaluations, all participants were shoeless and wore as little clothing as possible. Laboratory parameters Blood samples were collected by a qualified professional using the peripheral venous puncture technique with the Vacutainer system (Becton Dickinson, Plymouth, UK) between 7 and 10 a.m. after a 12-hour fast. The material was then centrifuged at 3000 rpm (2000 g) for 15 minutes. Serum 25(OH)D levels were evaluated by chemiluminescence (Alves et al., 2013) with an Architect 25(OH)D kit, using Architect/Abbott equipment (Abbott Architect I Instrument, IL, USA). 25(OH)D status was determined according to the following reference values: sufficient (≥ 30 ng/mL or 75 nmol/L), insufficient (≥ 20 ng/mL or 50 nmol/L to < 30 ng/mL or < 75 nmol/L), and deficient (< 20 ng/mL or < 50 nmol/L) [ 18 , 19 ]. Parathyroid hormone (PTH) was evaluated by chemiluminescence using the Access PTH Kit with UNICEL DXI800 equipment. Serum ultra-sensitive C-reactive protein (us-CRP) was determined by immunoturbidimetric assay (Bioclin, Quimbasa Quimica Básica, BH, MG, Brazil). Serum samples were frozen at 80°C for subsequent evaluation of interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70 and tumor necrosis factor (TNF)-α. The analysis was performed using the CBA human inflammatory cytokines kit (BD Biosciences, USA) according to the manufacturer's instructions, and the reading was performed on flow cytometry Facs Verse. Statistical analysis Data were tabulated in blinded duplicate using Epidata software. All statistical analyses were performed with the STATA statistical package, version 17.0. Normality of all variables was assessed using the Shapiro-Wilk test, skewness coefficient, and graphical analysis. Descriptive analysis was expressed as mean and 95% confidence intervals (CI) for continuous variables. Prevalence and 95% CI were recorded for categorical variables. After descriptive analysis, 25(OH)D, IL-1β, IL-6, IL-8, IL-10, IL-12p70, TNF-α, and us-CRP variables were log-transformed to improve normality. Differences in cytokines between 25(OH)D statuses were assessed using the test for trend across ordered groups. Multiple linear regression models (crude and adjusted) were used to determine associations between 25(OH)D concentration and inflammatory biomarkers. The models were adjusted for potential confounders: age group and sex (Model 1); PAL, season, cigarette smoking (current smoker, ex-smoker and never-smoker), BMI, schooling and PTH (Model 2). To test the interaction between serum 25(OH)D concentration and PAL categories, an interaction term (the product of PAL * 25(OH)D) was tested in the model, including the confounding variables. We also tested the interaction between 25(OH)D concentration and BMI in the adjusted model. All tests used a significance level of P < 0.05 and were performed with STATA version 17.0 (StataCorp, College Station, TX, USA). Results This study included data from 712 participants, 55.76% of whom were women. The mean age was 35.47 years, 24.8% of the sample were physically active, and the mean BMI was 25.12 (kg/m 2 ); other characteristics of the study population are shown in Table 1. In total, 11.6% of the population evaluated was considered deficient in 25(OH)D, 44.7% was insufficient, and 43.9% was sufficient (data not shown). Table 1 : Characteristics of study participants. Health and Food Study (ESA–Viçosa), 2012–2014. Parameter All (n=712) Age (years), mean (95% CI) Age group (years) 20-39 40-59 Sex (F) , % (95% CI) Schooling (years), mean (95% CI) 25(OH)D (ng/mL), mean (95% CI) BMI (kg/m2), mean (95% CI) PTH (pmol/L), mean (95% CI) IL-10 (pg/mL), median (95% CI) IL-6 (pg/mL), median (95% CI) IL-8 (pg/mL), median (95% CI) TNF-α (pg/mL), median (95% CI) IL-12p70 (pg/mL), median (95% CI) IL-1 (pg/mL), median (95% CI) us- CRP (mg/L), median (95% CI) PAL % (95% CI) <150 (minutes/week) ≥150 (minutes/week) Smoking status Never smoker Current smoker Former smoker Season % (95% CI) Summer/spring/fall Winter 35.47 (34.61; 36.33) 66.46 (63.45;69.34) 33.53 (30.65; 36.54) 55.76 (52.14; 59.32) 11.87 (11.55; 12.20) 30.17 (29.33; 31.02) 25.12 (24.76; 25.47) 26.85 (25.76; 27.95) 8.53 (7.94; 8.98) 11.81 (10.30; 12.98) 24.22 (23.04; 25.40) 8.28 (7.63; 9.46) 15.08 (14.05; 16.14) 7.40 (6.88; 8.10) 1.07 (0.96; 1.19) 75.2 (72.4; 77.8) 24.8 (22.2; 27.6) 70.61 (67.66; 73.40) 13.29 (11.30; 15.58) 16.08 (13.90;18.53) 86.61 (83.69; 89.07) 13.38 (10.92; 16.30) 25(OH)D: 25-hidroxivitamin D; BMI: body mass index; PTH: parathyroid hormone; IL: interleukin; TNF: tumor necrosis factor; us-CRP: ultra-sensitive C-reactive protein; PAL: physical activity level. Conversely, inflammatory biomarkers such as us-CRP (p<0.01) and IL-6 (p=0.03, Table 2) and serum PTH (Fig. 2A) were inversely associated with serum 25(OH)D status. A lower serum 25(OH)D concentration was observed in winter (25.49; 95%CI 23.89; 27.19 mmol/L) compared to other seasons (30.90; 95%CI 30.08; 31.72 mmol/L) (Fig. 2B). Table 2: Inflammatory biomarkers according to 25(OH)D status. Health and Food Study (ESA–Viçosa), 2012–2014. Biomarkers # Serum 25(OH)D status Deficient (< 50 nmol/L) Insufficient (≥ 50 and < 75 nmol/L) Sufficient (≥ 75 nmol/L) IL-6 (pg/mL) IL-10 (pg/mL) TNF-α (pg/mL) IL-12p70 (pg/mL) IL-8 (pg/mL) IL-1β (pg/mL) us-CRP (mg/L) 2.60 (2.43;2.77) 2.23 (2.11;2.37) 2.33 (2.12;2.52) 2.83 (2.64;3.01) 3.28 (3.15;3.40) 2.22 (2.04;2.40) 0.37 (0.10;0.60) 2.39 (2.30;2.48) 2.20 (2.14;2.27) 2.22 (2.13;2.31) 2.75 (2.66;2.84) 3.24 (3.18;3.31) 2.09 (2.00;2.18) 0.29 (0.14;0.55) 2.30 (2.18;2.41) 2.15 (2.13;2.27) 2.17 (2.06;2.27) 2.71 (2.61;2.81) 3.21 (3.14;3.28) 2.05 (1.95;2.15) -0.17 (-0.34;0.01) # Inflammatory biomarkers log transformed IL: interleukin; TNF: tumor necrosis factor; us-CRP: ultra-sensitive C-reactive protein. Fig. 3 shows the mean values of inflammatory biomarkers and 25(OH)D concentrations according to PAL. PAL was considered an effect modifier of the association between serum 25(OH)D levels and inflammatory biomarkers. Physically active (PA) individuals generally have lower mean concentrations of inflammatory biomarkers and higher 25(OH)D concentrations. Comparing IA with PA, only the differences in IL-6 [(IA: 2.38 - 95%CI 2.32; 2.44 pg/mL) vs. (PA: 2.19 - 95%CI 2.09; 2.31 pg/mL)] and 25(OH)D concentrations [(IA: 29.58 - 95%CI 28.76; 30.39 mg/mL) vs. (PA: 32.20 - 95%CI 30.53; 33.86 mg/mL)] were statistically significant. In the multiple linear regression models, it can be seen that for each unit increase in 25(OH)D concentration, there was an average decrease in the inflammatory biomarkers IL-6 (β= -0.37, 95%CI: -0.63; -0.10 pg/mL); IL-10 (β= -0.21, 95%CI: -0.37; -0.05 pg/mL); TNF-α (β=-0.29, 95%CI: -0.49; -0.07 pg/mL); IL-1β (β=-0.24, 95%CI: -0.45; -0.04 pg/mL); IL-12p70 (β= -0.22, 95%CI: -0.42; -0.02 pg/mL) and us-CRP (β= -0.46, 95%CI: -0.79; -0.12 pg/mL) in a model adjusted for age group and sex (Model 1). When we additionally adjusted for PAL, season, cigarette smoking, BMI, schooling and PTH, we found a negative association between 25(OH)D concentration and the inflammatory markers IL-6 (β= -0.29, 95%CI: -0.24; -0.07 pg/mL); IL-10 (β= -0.30, 95%CI: -0.57; -0.07 pg/mL); TNF-α (β=-0.30, 95%CI: -0.53; -0.07 pg/mL); IL-1β (β=-0.25, 95%CI: -0.47; -0.02 pg/mL) and IL-12p70 (β= -0.22, 95%CI: -0.44; -0.007 pg/mL) (Table 3). Moreover, PAL was considered an effect modifier of the association between serum 25(OH)D levels and IL-6 (p =0.03), IL-10 (p =0.008), IL-12p70 (p =0.014) and TNF-α (p = 0.018) levels. Therefore, association analyses were performed separately in IA and PA (Fig. 3). In participants with a PAL greater than 150 minutes, IL-6, IL-10, TNF-α and IL-12p70 levels tended to be lower as plasma 25(OH)D concentration increased (Fig. 4). Table 3 : Coefficients of linear regression models for the association between 25(OH)D status and inflammatory markers in the total population and according to physical activity level. Health and Food Study (ESA - Viçosa), 2012-2014 Inflammatory markers IL-6 (pg/mL)# β (CI95%) IL-10 (pg/mL)# β (CI95%) TNF-α (pg/mL)# β (CI95%) IL-8 (pg/mL)# β (CI95%) IL-1β (pg/mL)# β (CI95%) IL-12p70 (pg/mL)# β (CI95%) us-CRP (mg/L) # β (CI95%) 25(OH)D# Model 1 Model 2 -0.37 (-0.63; -0.10) -0.29 (-0.52; -0.06) -0.21(-0.37;-0.05) -0.24(-0.41;-0.07) -0.29 (-0.49; -0.07) -0.30 (-0.53; -0.07) -0.06 (-0.21; 0.09) -0.01 (-0.17; 0.15) -0.24 (-0.45;-0.04) -0.25 (-0.47;-0.02) -0.22 (-0.42; -0.02) -0.22 (-0.44; -0.007) -0.46 (-0.79;-0.12) -0.12 (-0.47; 0.22) Interaction 25(OH)D* PAL -0.44 (-0.72; -0.17) -0.38(-0.58; -0.1) -0.46 (-0.73; -0.18) -0.07(-0.26; 0.11) -0.46(-0.73;-0.18)* -0.34(-0.59; -0.06) 0.03(-0.38; 0.43) IL: interleukin; BMI: body mass index; PAL: physical activity level; TNF: tumor necrosis factor. # log transformed. Model 1: Adjusted for age group and sex. Model 2: Adjusted for age group, sex, season, PAL, smoking, BMI, schooling and PTH. * PAL variable and interaction term were not significant in model 2 . (p=0.321) Discussion The present study aimed to investigate the associations between 25(OH)D concentrations and inflammatory biomarkers in a Brazilian sample, and the presence of an interaction with physical activity and its mediating effect. We observed a negative association between 25(OH)D status and IL-6, IL-10, IL-1β, IL-12p70, and TNF-α in both models and us-CRP in the unadjusted model. This negative association remains in the presence of an interaction factor with PAL, confirming our hypothesis that higher 25(OH)D concentrations are associated with lower levels of inflammatory biomarkers in physically active adults. Elevated levels of inflammatory markers are associated with the pathogenesis of various metabolic disorders and non-communicable diseases. For instance, TNF-α and IL-1β play a role in the development of insulin resistance[ 20 ]; IL-6 has been linked to risk factors for cardiovascular diseases[ 21 ]; IL-12p70 has been identified as a critical marker in the early stages of atherosclerosis[ 22 ]; and CRP levels above 10 mg/L indicate a 4% increased risk of developing cardiovascular diseases[ 23 ]. This underscores the importance of understanding the factors that modulate inflammation in order to prevent and manage inflammation-related disorders and improve overall health outcomes. In general, vitamin D is known for its multiple roles in the body, including regulation of the immune system and modulation of inflammation. The multiple roles of vitamin D in the immune system are well studied, especially the immunomodulatory effect on decreasing proinflammatory cytokine production, immunoglobulin release, and increasing anti-inflammatory cytokine secretion[ 24 – 26 ]. In our study, us-CRP was inversely associated with serum 25(OH)D status, and for each unit increase in 25(OH)D concentration, an average decrease in the inflammatory biomarkers IL-6, IL-10, TNF-α, IL-1β, and IL-12p70 was observed. The mechanism by which 25(OH)D affects inflammatory markers is still unclear. However, there is evidence that 25(OH)D can modulate the production of inflammatory cytokines both in vivo and in vitro, decreasing the levels of inflammatory cytokines and increasing the levels of anti-inflammatory markers [ 27 – 29 ]. In addition, a systematic review found that 25(OH)D reduces the production of pro-inflammatory cytokines, including IL-6 and TNF-α[ 30 ]. There is some evidence to suggest that vitamin D deficiency may be associated with higher levels of inflammatory markers such as CRP and pro-inflammatory cytokines. However, the high prevalence of hypovitaminosis D observed reinforces that it is a global issue. The effectiveness of endogenous vitamin D (VD) production depends on environmental factors related to latitude, season and time of day of sun exposure, topical sunscreen use and skin pigmentation[ 6 ]. In the present sample, even though the city of Viçosa has a tropical climate, with mild winters and summers with abundant sunlight, a high prevalence of insufficient (44.7%) and deficient (11.6%) 25(OH)D levels was observed, with the lowest average observed in winter (25.49; 95%CI 23.89), compared to the other seasons. This is probably because in this season there is a lower incidence of UVB radiation and an increase in the use of clothing, consequently reducing the endogenous synthesis of vitamin D[ 31 ]. Higher PAL have been associated with adequate 25(OH)D status through several plausible mechanisms. First, individuals who engage in regular exercise, particularly outdoor activities, tend to have greater sun exposure, thereby increasing cutaneous 25(OH)D synthesis under adequate ultraviolet B (UVB) radiation[ 32 ]. Second, exercise promotes lipolysis in adipose tissue, potentially mobilizing vitamin D stored in fat and increasing circulating 25(OH)D concentrations[ 11 ]. Additionally, physically active individuals often maintain healthier body composition and metabolic profiles, which may optimize vitamin D metabolism and utilization[ 13 ]. Finally, an active lifestyle is generally associated with other healthy habits, such as a balanced diet and lower rates of obesity, which in turn contribute to improved vitamin D status[ 33 ]. In our study, being physically active (PA) was associated with lower levels of IL-6. This inflammatory marker can act as both pro- and anti-inflammatory. Its ambiguous role depends on the stimulus that triggers its secretion, being an anti-inflammatory marker when secreted by skeletal muscle[ 34 ]. A physically active lifestyle has been shown to lead to beneficial anti-inflammatory modulation. In general, the most sedentary group had higher odds of an elevated inflammatory status compared to the most active group[ 35 ]. Thus, it appears that physical activity may help to increase 25(OH)D levels and reduce inflammation, thereby promoting an anti-inflammatory state. Panagiotakos et al.[ 36 ] and Scragg et al.[ 32 ] have shown that high levels of physical activity are associated with attenuated circulating levels of inflammatory cytokines and that highly active individuals have significantly increased circulating levels of 25(OH)D compared to inactive individuals, which is consistent with our findings. However, there is controversy in the literature regarding the modulating effect of vitamin D, inflammatory markers and their mediation with physical activity levels, possibly due to methodological issues. Carrillo et al. (2012)[ 7 ] observed that 12 weeks of resistance training with (n = 13, 53.8% women) or without (n = 10, 50.0% women) vitamin D supplementation (4000 IU/day) did not improve IL-6, CRP or TNF-α. However, the sample was recruited in winter and the authors provided and instructed the sample to use sunscreen every day, which could have interfered with the effect of outdoor physical activity on 25(OH)D levels. Moreover, in addition to the resistance training intervention, the sample (BMI > 25 kg/m 2 ) was weight stable, which could mean that the mobilization of vitamin D stored in the body was impaired. In contrast, Nazarabadi et al. (2022)[ 37 ] observed that in a sample of 46 postmenopausal women with metabolic syndrome, the intervention of physical activity or vitamin D supplementation (50000 IU/day) for 8 weeks decreased CRP and IL-6 levels and improved metabolic syndrome parameters. And the authors observed that the physical activity plus vitamin D supplementation group produced better results than physical activity or vitamin D supplementation alone. To understand the modulating effect of vitamin D, inflammatory markers and their mediation with the level of physical activity, an interaction term was included in the adjusted model in our study. As a result, we observed that the mean levels of inflammatory markers decreased in individuals who had higher levels of PAL and 25(OH)D concentration. Therefore, it seems that while vitamin D deficiency is associated with greater inflammation, physical activity tends to have a beneficial effect on both, potentially helping to increase vitamin D levels and reduce inflammatory markers. The present study had some limitations, such as its cross-sectional design, which did not allow inferences about the direction of the associations found. However, the cluster sampling and the population-based design mitigated this limitation. Although the IPAQ is widely used and standardized, it relies on self-reported information, which is subject to recall bias, over- or underestimation of activity duration and intensity, and social desirability bias. Furthermore, the IPAQ does not capture detailed temporal patterns of physical activity or account for activities of short duration but high intensity. For future studies aiming to obtain more objective and precise measures, direct assessment methods or other wearable devices are recommended. These devices can provide real-time data on activity intensity, frequency, and duration, thereby reducing measurement error and allowing for more accurate classification of PAL. Strengths of this study include the population-based sample size of 712 individuals and the methodological rigor applied in the data collection process, such as training the entire team involved and conducting a pilot study. These factors reinforced the quality and ensured the validity of the results obtained. Conclusion 25(OH)D and physical activity play complementary roles in reducing inflammation. Controlling vitamin D deficiency and promoting an active lifestyle, preferably outdoors, may be critical strategies to improve population health by reducing the impact of elevated levels of inflammatory markers, potentially contributing to overall well-being. Declarations Ethics approval and consent to participate The research project was approved by the Research Ethics Committee of the Federal University of Viçosa (Official Letter 02/2013). Consent for publication All participants signed the informed consent form. Availability of data and materials The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing interests None declared. Funding This research received financial support from the National Council for Scientific and Technological Development (CNPq process 481418/2011 3), the Research Support Foundation of the State of Minas Gerais (FAPEMIG process APQ-00296 12). Authors' contributions Giana Zarbato Longo: Conceived the study idea, was responsible for statistical analysis and coordinator of the study. Danielle Cristina Guimarães da Silva: data acquisition and wrote the main text. Mariana Papinni Gabiatti: made many useful suggestions during the revision of the manuscript. Fernanda Hansen: made critical comments and revisions for the initial manuscript. Fabrícia Geralda Ferreira: wrote the initial manuscript. Diego Augusto Santos Silva: physical education expertise and significant reviewer. Leandro Licursi de Oliveira: performed the biochemical analysis of cytokines. Amanda Alcaraz da Silva: wrote the discussion text. Yara Maria Franco Moreno: made critical comments and revisions for the initial manuscript. Acknowledgment We would like to thank all of the participants for their dedication National Council for Scientific and Technological Development and also the Foundations for Supporting Research in the states of Minas Gerais for financially funding the project. The federal university of viçosa and its facilities for the study. References Holick MF, Chen TC. Vitamin D deficiency: A worldwide problem with health consequences. 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Int J Vitam Nutr Res 2017;87:49–58. https://doi.org/10.1024/0300-9831/a000424. Sun X, Cao ZB, Taniguchi H, Tanisawa K, Higuchi M. Effect of an acute bout of endurance exercise on serum 25(OH)D concentrations in young adults. J Clin Endocrinol Metab 2017;102:3937–44. https://doi.org/10.1210/jc.2017-00146. Pannu PK, Zhao Y, Soares MJ. Reductions in body weight and percent fat mass increase the vitamin D status of obese subjects: A systematic review and metaregression analysis. Nutr Res 2016;36:201–13. https://doi.org/10.1016/j.nutres.2015.11.013. Simpson RJ, Krüger K, Walsh NP, Campbell JP, Gleeson M, Nieman DC, et al. Can exercise affect immune function to increase susceptibility to infection? Exerc Immunol Rev 2020:8–22. Wang J, Liu S, Li G, Xiao J. Exercise Regulates the Immune System. Adv Exp Med Biol 2020;1228:395–408. https://doi.org/10.1007/978-981-15-1792-1_27. Segheto W, Cristina Guimarães da Silva D, Araújo Coelho F, Guimarães Reis V, Helena Oliveira Morais S, Carlos Bouzas Marins J, et al. Body Adiposity Index and Associated Factors in Adults: Method and Logistics of a Population-Based Study. Nutr Hosp 2015;32:101–9. https://doi.org/10.3305/nh.2015.32.1.8391. Steele EM, Claro RM, Monteiro CA. Behavioural patterns of protective and risk factors for non-communicable diseases in Brazil. Public Health Nutr 2014;17:369–75. https://doi.org/10.1017/S1368980012005472. World Health Organization. WHO guidelines on physical activity and sedentary behaviour: at a glance. World Heal Organ 2020:535. Matsudo S, Araújo T, Matsudo V, Andrade D, Andrade E, Oliveira LC, et al. Questionário Internacional De Atividade Física (Ipaq): Estupo De Validade E Reprodutibilidade No Brasil. Rev Bras Atividade Física Saúde 2012;6:5–18. https://doi.org/10.12820/rbafs.v.6n2p5-18. Pludowski P, Takacs I, Boyanov M, Belaya Z, Diaconu CC, Mokhort T, et al. Clinical Practice in the Prevention, Diagnosis and Treatment of Vitamin D Deficiency: A Central and Eastern European Expert Consensus Statement. Nutrients 2022;14:1–18. https://doi.org/10.3390/nu14071483. Holick MF. Resurrection of vitamin D deficiency and rickets. J Clin Invest 2006;116:2062–72. https://doi.org/10.1172/JCI29449. Thorsen SU, Pipper CB, Skogstrand K, Pociot F, Svensson J. 25-hydroxyvitamin D and peripheral immune mediators: Results from two nationwide danish pediatric cohorts. Nutrients 2017;9:4–13. https://doi.org/10.3390/nu9040365. Haddy N, Sass C, Droesch S, Zaiou M, Siest G, Ponthieux A, et al. IL-6, TNF-α and atherosclerosis risk indicators in a healthy family population: The STANISLAS cohort. Atherosclerosis 2003;170:277–83. https://doi.org/10.1016/S0021-9150(03)00287-9. Yong K, Dogra G, Boudville N, Chan D, Adams L, Ching H, et al. Interleukin-12 is associated with arterial stiffness in healthy individuals. Am J Hypertens 2013;26:159–62. https://doi.org/10.1093/ajh/hps032. Cozlea DL, Farcas DM, Nagy A, Keresztesi AA, Tifrea R, Cozlea L, et al. The impact of C reactive protein on global cardiovascular risk on patients with coronary artery disease. Curr Heal Sci J 2013;39:225–31. Lemire JM. lmmunomodulatory Role of 1 , 25-Dihydroxyvitamin D3 1984;31:26–31. Zhang Y, Leung DYM, Richers BN, Liu Y, Remigio LK, Riches DW, et al. Vitamin D Inhibits Monocyte/Macrophage Proinflammatory Cytokine Production by Targeting MAPK Phosphatase-1. J Immunol 2012;188:2127–35. https://doi.org/10.4049/jimmunol.1102412. Chen S, Lipsky PE, Chen S, Sims GP, Chen XX, Gu YY. on Human B Cell Differentiation 3 D Modulatory Effects of 1,25-Dihydroxyvitamin Modulatory Effects of 1,25-Dihydroxyvitamin D 3 on Human B Cell Differentiation. J Immunol Ref J Immunol Samsung Med Cent 2007;179:1634–47. Akbari M, Ostadmohammadi V, Lankarani KB, Tabrizi R, Kolahdooz F, Heydari ST, et al. The Effects of Vitamin D Supplementation on Biomarkers of Inflammation and Oxidative Stress among Women with Polycystic Ovary Syndrome: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. Horm Metab Res 2018;50:271–9. https://doi.org/10.1055/s-0044-101355. Mansournia MA, Ostadmohammadi V, Doosti-Irani A, Ghayour-Mobarhan M, Ferns G, Akbari H, et al. Abbreviations GSH Total glutathione hs-CRP High-sensitivity C-reactive protein MDA Malondialdehyde NO Nitric oxide TAC Total antioxidant capacity The Effects of Vitamin D Supplementation on Biomarkers of Inflammation and Oxidative Stress in Diabetic Patie. Vitam D Diabetes … Horm Metab Res Horm Metab Res Mansournia MA Al Vitam D Diabetes … Horm Metab Res 2018;50:429–40. Sepidarkish M, Farsi F, Akbari-Fakhrabadi M, Namazi N, Almasi-Hashiani A, Maleki Hagiagha A, et al. The effect of vitamin D supplementation on oxidative stress parameters: A systematic review and meta-analysis of clinical trials. Pharmacol Res 2019;139:141–52. https://doi.org/10.1016/j.phrs.2018.11.011. Saedmocheshi S, Amiri E, Mehdipour A, Stefani GP. The Effect of Vitamin D Consumption on Pro-Inflammatory Cytokines in Athletes: A Systematic Review of Randomized Controlled Trials. Sports 2024;12. https://doi.org/10.3390/sports12010032. Holick MF. Vitamin D: A D-lightful solution for health. J Investig Med 2011;59:872–80. https://doi.org/10.2310/JIM.0b013e318214ea2d. Scragg R, Camargo CA. Frequency of leisure-time physical activity and serum 25-hydroxyvitamin D levels in the US population: Results from the third national health and nutrition examination survey. Am J Epidemiol 2008;168:577–86. https://doi.org/10.1093/aje/kwn163. Segheto KJ, Pereira M, Da Silva DCG, De Carvalho CJ, Massardi FR, Kakehasi AM, et al. Vitamin d and bone health in adults: A systematic review and meta-analysis. Cienc e Saude Coletiva 2021;26:3221–44. https://doi.org/10.1590/1413-81232021268.15012020. Villar-Fincheira P, Sanhueza-Olivares F, Norambuena-Soto I, Cancino-Arenas N, Hernandez-Vargas F, Troncoso R, et al. Role of Interleukin-6 in Vascular Health and Disease. Front Mol Biosci 2021;8:1–11. https://doi.org/10.3389/fmolb.2021.641734. Cho SMJ, Lee H, Shim JS, Jeon JY KH. Association between physical activity and inflammatory markers in community-dwelling, middle-aged adults. Appl Physiol Nutr Metab 2021;33566730:1–31. https://doi.org/10.1139/apnm-2020-1069. Panagiotakos DB, Pitsavos C, Chrysohoou C, Kavouras S, Stefanadis C. The associations between leisure-time physical activity and inflammatory and coagulation markers related to cardiovascular disease: The ATTICA Study. Prev Med (Baltim) 2005;40:432–7. https://doi.org/10.1016/j.ypmed.2004.07.010. Nazarabadi PN, Etemad Z, Hoseini R MF. Anti-Inflammatory Effects of a Period of Aerobic Training and Vitamin D Supplementation in Postmenopausal Women with Metabolic Syndrome. Int J Prev Med 2022;13:1–7. 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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-6214919","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":430260220,"identity":"54a365c7-8c5d-479d-b627-0d8f1a93ad0f","order_by":0,"name":"Giana Zarbato Longo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/ElEQVRIiWNgGAWjYDACZhhDAkRUgESYG4jWwtjAcAYkwkhACxyAtDC2gVgEtMi7Mz978IOhTt58dvPzBx/n1UbztwO1/KjYhlOL4WE2c8MeBjbDOXeOGTbO3HY8d8ZhoG09Z27j1tLMYCbBw8DDOEMiwbCZd9ux3AagFmbGNnxa2L9J/mGQsJ8hkf6xmXfOsdz5hLTIM/OYSfMwGCTOkMgB2tJQk7uBkBYDZp4yaRmDhOQZMmcKZ844diB3I1DLQXx+ke8/vk3yTUWd7Qzp9g0fPtTU5c47f/jggx8VeGw5ACbh/MNg8gBO9SBbGlD5dfgUj4JRMApGwQgFACrGVqU5CxDbAAAAAElFTkSuQmCC","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":true,"prefix":"","firstName":"Giana","middleName":"Zarbato","lastName":"Longo","suffix":""},{"id":430260227,"identity":"5b3dfc1e-b5cd-4ae6-afae-6a53ec7b8c07","order_by":1,"name":"Danielle Cristina Guimarães da Silva","email":"","orcid":"","institution":"Federal University of Western Bahia","correspondingAuthor":false,"prefix":"","firstName":"Danielle","middleName":"Cristina Guimarães da","lastName":"Silva","suffix":""},{"id":430260230,"identity":"35d1e08b-957e-450a-b1cb-0cc03ed1211b","order_by":2,"name":"Mariana Papinni Gabiatti","email":"","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Mariana","middleName":"Papinni","lastName":"Gabiatti","suffix":""},{"id":430260232,"identity":"c4b7edf6-90f3-4187-a190-95ccfc01fd9f","order_by":3,"name":"Fernanda Hansen","email":"","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Fernanda","middleName":"","lastName":"Hansen","suffix":""},{"id":430260235,"identity":"6034da7f-272e-4c1c-b706-a0bf5fb96e85","order_by":4,"name":"Fabricia Geralda Ferreira","email":"","orcid":"","institution":"Universidade da Força Aérea","correspondingAuthor":false,"prefix":"","firstName":"Fabricia","middleName":"Geralda","lastName":"Ferreira","suffix":""},{"id":430260237,"identity":"7fea3217-544f-4749-a71a-71d351340f8d","order_by":5,"name":"Diego Augusto Santos Silva","email":"","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Diego","middleName":"Augusto Santos","lastName":"Silva","suffix":""},{"id":430260238,"identity":"70640319-8a6d-425e-8393-c89bdfd69ed8","order_by":6,"name":"Leandro Licursi de Oliveira","email":"","orcid":"","institution":"Universidade Federal de Viçosa","correspondingAuthor":false,"prefix":"","firstName":"Leandro","middleName":"Licursi","lastName":"de Oliveira","suffix":""},{"id":430260240,"identity":"6288644a-aca0-453d-81ea-be615bb229a8","order_by":7,"name":"Amanda Alcaraz da Silva","email":"","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Amanda","middleName":"Alcaraz da","lastName":"Silva","suffix":""},{"id":430260243,"identity":"5b93a169-d2bf-4ff3-9093-b5a9c77554b9","order_by":8,"name":"Yara Maria Franco Moreno","email":"","orcid":"","institution":"Universidade Federal de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Yara","middleName":"Maria Franco","lastName":"Moreno","suffix":""}],"badges":[],"createdAt":"2025-03-12 21:23:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6214919/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6214919/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":78884811,"identity":"1eb4d1f0-3b23-4c47-9db9-4082f471d790","added_by":"auto","created_at":"2025-03-20 09:24:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":22530,"visible":true,"origin":"","legend":"\u003cp\u003eFlowchart of participant selection. City of Viçosa, State of Minas Gerais, Brazil.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6214919/v1/b1b1ef3b19503cc2bebdb962.png"},{"id":78886735,"identity":"ab4c68fd-2ca5-4e84-a3ba-4f0de084ffeb","added_by":"auto","created_at":"2025-03-20 09:48:59","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":46423,"visible":true,"origin":"","legend":"\u003cp\u003eDifferences between 25(OH)D concentrations with serum PTH (A) and season (B) in Brazilian population. Health and Food Study (ESA–Viçosa), 2012–2014.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6214919/v1/efe1a497eb904f7ac1062a1e.png"},{"id":78884814,"identity":"acd8d376-89ad-45fa-be9a-da85cc1a1603","added_by":"auto","created_at":"2025-03-20 09:24:59","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":99975,"visible":true,"origin":"","legend":"\u003cp\u003eMean values of inflammatory biomarkers and 25(OH)D concentrations according to physical activity level (PAL). Health and Food Study (ESA–Viçosa), 2012–2014.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6214919/v1/3a41a24e0e4cc1f8c35b3672.png"},{"id":78885263,"identity":"15ba4c6a-1a76-44e8-9962-004ddda25533","added_by":"auto","created_at":"2025-03-20 09:32:59","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":287391,"visible":true,"origin":"","legend":"\u003cp\u003eAssociation between inflammatory biomarkers, PAL, and 25(OH)D levels in a Brazilian population. Black circle symbols and gray triangle symbols represent observed values of PAL \u0026lt; 150 minutes and PAL \u0026gt;= 150 minutes, respectively. Lines represent coefficients of 25(OH)D obtained by multiple regression adjusted for sex, age group, season, PAL, smoking, sedentary time, BMI, schooling, and PTH.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6214919/v1/0ddab4ea433e065749a5bda1.png"},{"id":79067007,"identity":"145bd6ee-d1ed-4d4e-b904-1c9bd6ee0f1b","added_by":"auto","created_at":"2025-03-24 04:38:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1226657,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6214919/v1/3187d5b9-f024-4afe-bdaf-d39b7409197c.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Association between Cytokines, 25-Hydroxyvitamin D, and Physical Activity: Evidence from a cross sectional Brazilian Population","fulltext":[{"header":"Background","content":"\u003cp\u003eVitamin D (25(OH)D) deficiency is a global problem and is prevalent in tropical countries[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. In Brazil, a meta-analysis aimed at studying the prevalence of deficiency and insufficiency of this vitamin in the Brazilian population in different age groups found values of 28.16% (95% CI: 23.90, 32.40) and 45.26% (95% CI: 35.82, 54.71)[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], respectively, confirming that hypovitaminosis D is a public health problem.\u003c/p\u003e \u003cp\u003eResearch conducted in recent years has shown the association between hypovitaminosis D and several diseases, including cancer[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], cardiometabolic disease[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], and inflammatory disorders[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], and has highlighted the association of this vitamin with some inflammatory cytokines and immune signaling, attracting increasing attention from the scientific community. 25(OH)D can reduce systemic inflammatory mediators and promote the release of anti-inflammatory cytokines from the immune system. A possible explanation for this relationship may be the anti-inflammatory properties of 25(OH)D, which affects the production and function of adipokines, thereby modulating the inflammatory response in adipose tissue[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. In the NHANES study, in a probabilistic sample of US adults, individuals with high serum concentrations of 25(OH)D were found to have a better profile of inflammatory biomarkers[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFew studies have investigated the relationship between the effect of 25(OH)D in combination with physical activity throughout the seasons[\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The interaction between physical activity and serum 25(OH)D levels has been explored by several studies, suggesting physiological mechanisms that explain this relationship and the potential effect on inflammation modulation. One of the main hypotheses is that physical activity stimulates lipolysis in adipose tissue, releasing the vitamin D stored there and consequently increasing serum 25(OH)D concentrations[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. In addition, regular physical activity is associated with an improved metabolic profile, reduced body adiposity, and maintenance of a healthy weight, factors that may also positively influence vitamin D status[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. From an immunological point of view, regular physical activity stimulates the production of anti-inflammatory myokines (e.g. IL-10) and reduces the concentration of pro-inflammatory cytokines (e.g. TNF-α), thus contributing to a more balanced immune environment[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Therefore, when combined, adequate levels of 25(OH)D and physical activity can potentiate the reduction of systemic inflammatory markers.\u003c/p\u003e \u003cp\u003eConsidering the possible modulating effect of 25(OH)D) on inflammation and its influence on the reduction of inflammatory markers associated with a sufficient level of physical activity, our hypothesis is that higher plasma levels of 25(OH)D are associated with lower plasma concentrations of cytokines in physically active individuals. Therefore, the aim of this study was to investigate the association between inflammatory biomarkers and 25(OH)D concentration in a sample of Brazilian adults, taking into account the influence of physical activity on this association.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy design and participants\u003c/h2\u003e \u003cp\u003eThis is a population-based cross-sectional study aimed at assessing the health status of the adult population of Vi\u0026ccedil;osa/MG, Brazil (2012\u0026ndash;2014). The sample was composed of non-institutionalized individuals of both sexes, between 20 and 59 years of age, who were residents of the urban area of the city.\u003c/p\u003e \u003cp\u003eThe sampling process was probabilistic without repositioning in two stages (census sectors and domicile). In the end, 1229 home interviews were conducted; 331 of the interviewees did not complete the laboratory examination and 186 were excluded from the sample due to loss of the biological sample. The final sample size was 712 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). There were no statistical differences in sociodemographic characteristics between the original sample and the sample used in this study (data not shown).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e This study was conducted in accordance with the Declaration of Helsinki. The research project was approved by the Research Ethics Committee of the Federal University of Vi\u0026ccedil;osa (Official Letter 02/2013). All participants signed the informed consent form. Additional methodological details can be found in Segheto et al. (2015)[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSociodemographic, behavioral and seasonality variables\u003c/h3\u003e\n\u003cp\u003eThe sociodemographic variables were: age, expressed as completed years and age group (20\u0026ndash;39; 40\u0026ndash;59 years); sex (male and female); years of schooling completed. The behavioral variables were: cigarette smoking (current smoker, ex-smoker and never-smoker) regardless of frequency and intensity of tobacco use[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]; and physical activity level (PAL). PAL was assessed using the long version (version 6) of the International Physical Activity Questionnaire (IPAQ) as described by Matsudo et al. (2001) [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. For this study, only time spent in leisure-time physical activity during the week prior to administration of the questionnaire was considered. Participants were then categorized as irregularly active (IA) (\u0026lt;\u0026thinsp;150 min/week) or physically active (PA) (\u0026ge;\u0026thinsp;150 min/week)[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] (World Health Organization, 2020).\u003c/p\u003e \u003cp\u003eThe seasonality variable, which indicates the time of year when blood was collected to assess vitamin D status, was dichotomized into winter and summer/spring/autumn.\u003c/p\u003e\n\u003ch3\u003eAnthropometric parameters\u003c/h3\u003e\n\u003cp\u003eBody mass index (BMI) was calculated as body mass (kg) divided by height (m) squared, expressed in kilograms per meter squared (kg/m\u003csup\u003e2\u003c/sup\u003e). Body mass was measured with a digital scale (Tanita, Ironman TM model, BC-554, Tanita Corporation), and height was measured using a metal stadiometer (Welmy, Santa B\u0026aacute;rbara D'Oeste, SP, Brazil). During the anthropometric evaluations, all participants were shoeless and wore as little clothing as possible.\u003c/p\u003e\n\u003ch3\u003eLaboratory parameters\u003c/h3\u003e\n\u003cp\u003eBlood samples were collected by a qualified professional using the peripheral venous puncture technique with the Vacutainer system (Becton Dickinson, Plymouth, UK) between 7 and 10 a.m. after a 12-hour fast. The material was then centrifuged at 3000 rpm (2000 g) for 15 minutes.\u003c/p\u003e \u003cp\u003eSerum 25(OH)D levels were evaluated by chemiluminescence (Alves et al., 2013) with an Architect 25(OH)D kit, using Architect/Abbott equipment (Abbott Architect I Instrument, IL, USA). 25(OH)D status was determined according to the following reference values: sufficient (\u0026ge;\u0026thinsp;30 ng/mL or 75 nmol/L), insufficient (\u0026ge;\u0026thinsp;20 ng/mL or 50 nmol/L to \u0026lt;\u0026thinsp;30 ng/mL or \u0026lt;\u0026thinsp;75 nmol/L), and deficient (\u0026lt;\u0026thinsp;20 ng/mL or \u0026lt;\u0026thinsp;50 nmol/L) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eParathyroid hormone (PTH) was evaluated by chemiluminescence using the Access PTH Kit with UNICEL DXI800 equipment. Serum ultra-sensitive C-reactive protein (us-CRP) was determined by immunoturbidimetric assay (Bioclin, Quimbasa Quimica B\u0026aacute;sica, BH, MG, Brazil).\u003c/p\u003e \u003cp\u003eSerum samples were frozen at 80\u0026deg;C for subsequent evaluation of interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70 and tumor necrosis factor (TNF)-α. The analysis was performed using the CBA human inflammatory cytokines kit (BD Biosciences, USA) according to the manufacturer's instructions, and the reading was performed on flow cytometry Facs Verse.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eData were tabulated in blinded duplicate using Epidata software. All statistical analyses were performed with the STATA statistical package, version 17.0. Normality of all variables was assessed using the Shapiro-Wilk test, skewness coefficient, and graphical analysis. Descriptive analysis was expressed as mean and 95% confidence intervals (CI) for continuous variables. Prevalence and 95% CI were recorded for categorical variables. After descriptive analysis, 25(OH)D, IL-1β, IL-6, IL-8, IL-10, IL-12p70, TNF-α, and us-CRP variables were log-transformed to improve normality.\u003c/p\u003e \u003cp\u003eDifferences in cytokines between 25(OH)D statuses were assessed using the test for trend across ordered groups.\u003c/p\u003e \u003cp\u003eMultiple linear regression models (crude and adjusted) were used to determine associations between 25(OH)D concentration and inflammatory biomarkers. The models were adjusted for potential confounders: age group and sex (Model 1); PAL, season, cigarette smoking (current smoker, ex-smoker and never-smoker), BMI, schooling and PTH (Model 2). To test the interaction between serum 25(OH)D concentration and PAL categories, an interaction term (the product of PAL * 25(OH)D) was tested in the model, including the confounding variables. We also tested the interaction between 25(OH)D concentration and BMI in the adjusted model. All tests used a significance level of P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and were performed with STATA version 17.0 (StataCorp, College Station, TX, USA).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eThis study included data from 712 participants, 55.76% of whom were women. The mean age was 35.47 years, 24.8% of the sample were physically active, and the mean BMI was 25.12 (kg/m\u003csup\u003e2\u003c/sup\u003e); other characteristics of the study population are shown in Table 1. In total, 11.6% of the population evaluated was considered deficient in 25(OH)D, 44.7% was insufficient, and 43.9% was sufficient (data not shown).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e: Characteristics of study participants. Health and Food Study (ESA\u0026ndash;Vi\u0026ccedil;osa), 2012\u0026ndash;2014.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"536\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 347px;\"\u003e\n \u003cp\u003eParameter\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\n \u003cp\u003eAll (n=712)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 347px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge\u003c/strong\u003e (years), mean (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eAge group\u0026nbsp;\u003c/strong\u003e(years)\u003c/p\u003e\n \u003cp\u003e20-39\u003c/p\u003e\n \u003cp\u003e40-59\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSex (F)\u003c/strong\u003e, % (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSchooling\u003c/strong\u003e (years), mean (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e25(OH)D\u0026nbsp;\u003c/strong\u003e (ng/mL), mean (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eBMI\u003c/strong\u003e (kg/m2), mean (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ePTH\u003c/strong\u003e (pmol/L), mean (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIL-10\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIL-6\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIL-8\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTNF-\u0026alpha;\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIL-12p70\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIL-1\u003c/strong\u003e (pg/mL), median (95% CI)\u003c/p\u003e\n \u003cp\u003eus-\u003cstrong\u003eCRP\u003c/strong\u003e (mg/L), median (95% CI)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ePAL\u003c/strong\u003e % (95% CI)\u003c/p\u003e\n \u003cp\u003e\u0026lt;150 (minutes/week)\u003c/p\u003e\n \u003cp\u003e\u0026ge;150 (minutes/week)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSmoking status\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eNever smoker\u003c/p\u003e\n \u003cp\u003eCurrent smoker\u003c/p\u003e\n \u003cp\u003eFormer smoker\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSeason\u0026nbsp;\u003c/strong\u003e% (95% CI)\u003c/p\u003e\n \u003cp\u003eSummer/spring/fall\u003c/p\u003e\n \u003cp\u003eWinter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\n \u003cp\u003e35.47 (34.61; 36.33)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e66.46 (63.45;69.34)\u003c/p\u003e\n \u003cp\u003e33.53 (30.65; 36.54)\u003c/p\u003e\n \u003cp\u003e55.76 (52.14; 59.32)\u003c/p\u003e\n \u003cp\u003e11.87 (11.55; 12.20)\u003c/p\u003e\n \u003cp\u003e30.17 (29.33; 31.02)\u003c/p\u003e\n \u003cp\u003e25.12 (24.76; 25.47)\u003c/p\u003e\n \u003cp\u003e26.85 (25.76; 27.95)\u003c/p\u003e\n \u003cp\u003e8.53 (7.94; 8.98)\u003c/p\u003e\n \u003cp\u003e11.81 (10.30; 12.98)\u003c/p\u003e\n \u003cp\u003e24.22 (23.04; 25.40)\u003c/p\u003e\n \u003cp\u003e8.28 (7.63; 9.46)\u003c/p\u003e\n \u003cp\u003e15.08 (14.05; 16.14)\u003c/p\u003e\n \u003cp\u003e7.40 (6.88; 8.10)\u003c/p\u003e\n \u003cp\u003e1.07 (0.96; 1.19)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e75.2 (72.4; 77.8)\u003c/p\u003e\n \u003cp\u003e24.8 (22.2; 27.6)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e70.61 (67.66; 73.40)\u003c/p\u003e\n \u003cp\u003e13.29 (11.30; 15.58)\u003c/p\u003e\n \u003cp\u003e16.08 (13.90;18.53)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e86.61 (83.69; 89.07)\u003c/p\u003e\n \u003cp\u003e13.38 (10.92; 16.30)\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e25(OH)D: 25-hidroxivitamin D; BMI: body mass index; PTH: parathyroid hormone; IL: interleukin; TNF:\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003etumor necrosis factor; us-CRP: ultra-sensitive C-reactive protein; PAL: physical activity level.\u003c/p\u003e\n\u003cp\u003eConversely, inflammatory biomarkers such as us-CRP (p\u0026lt;0.01) and IL-6 (p=0.03, Table 2) and serum PTH (Fig. 2A) were inversely associated with serum 25(OH)D status. A lower serum 25(OH)D concentration was observed in winter (25.49; 95%CI 23.89; 27.19 mmol/L) compared to other seasons (30.90; 95%CI 30.08; 31.72 mmol/L) (Fig. 2B).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2:\u003c/strong\u003e Inflammatory biomarkers according to 25(OH)D status. Health and Food Study (ESA\u0026ndash;Vi\u0026ccedil;osa), 2012\u0026ndash;2014.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"568\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eBiomarkers #\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 425px;\"\u003e\n \u003cp\u003eSerum 25(OH)D status\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 141px;\"\u003e\n \u003cp\u003eDeficient\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;(\u0026lt; 50 nmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003eInsufficient\u003c/p\u003e\n \u003cp\u003e(\u0026ge; 50 and\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u0026lt; 75 nmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 139px;\"\u003e\n \u003cp\u003eSufficient\u003c/p\u003e\n \u003cp\u003e(\u0026ge; 75 nmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eIL-6 (pg/mL)\u003c/p\u003e\n \u003cp\u003eIL-10 (pg/mL)\u003c/p\u003e\n \u003cp\u003eTNF-\u0026alpha;\u0026nbsp;(pg/mL)\u003c/p\u003e\n \u003cp\u003eIL-12p70 (pg/mL)\u003c/p\u003e\n \u003cp\u003eIL-8 (pg/mL)\u003c/p\u003e\n \u003cp\u003eIL-1\u0026beta;\u0026nbsp;(pg/mL)\u003c/p\u003e\n \u003cp\u003eus-CRP (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.60 (2.43;2.77)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e2.23 (2.11;2.37)\u003c/p\u003e\n \u003cp\u003e2.33 (2.12;2.52)\u003c/p\u003e\n \u003cp\u003e2.83 (2.64;3.01)\u003c/p\u003e\n \u003cp\u003e3.28 (3.15;3.40)\u003c/p\u003e\n \u003cp\u003e2.22 (2.04;2.40)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.37 (0.10;0.60)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 145px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.39 (2.30;2.48)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e2.20 (2.14;2.27)\u003c/p\u003e\n \u003cp\u003e2.22 (2.13;2.31)\u003c/p\u003e\n \u003cp\u003e2.75 (2.66;2.84)\u003c/p\u003e\n \u003cp\u003e3.24 (3.18;3.31)\u003c/p\u003e\n \u003cp\u003e2.09 (2.00;2.18)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.29 (0.14;0.55)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 139px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.30 (2.18;2.41)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e2.15 (2.13;2.27)\u003c/p\u003e\n \u003cp\u003e2.17 (2.06;2.27)\u003c/p\u003e\n \u003cp\u003e2.71 (2.61;2.81)\u003c/p\u003e\n \u003cp\u003e3.21 (3.14;3.28)\u003c/p\u003e\n \u003cp\u003e2.05 (1.95;2.15)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.17 (-0.34;0.01)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e# Inflammatory biomarkers log transformed\u003c/p\u003e\n\u003cp\u003eIL: interleukin; TNF:\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003etumor necrosis factor; us-CRP: ultra-sensitive C-reactive protein.\u003c/p\u003e\n\u003cp\u003eFig. 3 shows the mean values of inflammatory biomarkers and 25(OH)D concentrations according to PAL. PAL was considered an effect modifier of the association between serum 25(OH)D levels and inflammatory biomarkers. Physically active (PA) individuals generally have lower mean \u0026nbsp;concentrations of inflammatory biomarkers and higher 25(OH)D concentrations. Comparing IA with PA, only the differences in IL-6 [(IA: \u0026nbsp; 2.38 - 95%CI 2.32; 2.44 pg/mL) vs. (PA: 2.19 - 95%CI 2.09; 2.31 pg/mL)] and 25(OH)D concentrations [(IA: \u0026nbsp; 29.58 - 95%CI 28.76; 30.39 mg/mL) vs. (PA: 32.20 - 95%CI 30.53; 33.86 mg/mL)] were statistically significant.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn the multiple linear regression models, it can be seen that for each unit increase in 25(OH)D concentration, there was an average decrease in the inflammatory biomarkers IL-6 (\u0026beta;= -0.37, 95%CI: -0.63; -0.10 pg/mL); IL-10 (\u0026beta;= -0.21, 95%CI: -0.37; -0.05 pg/mL); TNF-\u0026alpha; (\u0026beta;=-0.29, 95%CI: -0.49; -0.07 pg/mL); IL-1\u0026beta; (\u0026beta;=-0.24, 95%CI: -0.45; -0.04 pg/mL); IL-12p70 (\u0026beta;= -0.22, 95%CI: -0.42; -0.02 pg/mL) and us-CRP (\u0026beta;= -0.46, 95%CI: -0.79; -0.12 pg/mL) in a model adjusted for age group and sex (Model 1). When we additionally adjusted for PAL, season, cigarette smoking, BMI, schooling and PTH, we found a negative association between 25(OH)D concentration and the inflammatory markers IL-6 (\u0026beta;= -0.29, 95%CI: -0.24; -0.07 pg/mL); IL-10 (\u0026beta;= -0.30, 95%CI: -0.57; -0.07 pg/mL); TNF-\u0026alpha; (\u0026beta;=-0.30, 95%CI: -0.53; -0.07 pg/mL); IL-1\u0026beta; (\u0026beta;=-0.25, 95%CI: -0.47; -0.02 pg/mL) and IL-12p70 (\u0026beta;= -0.22, 95%CI: -0.44; -0.007 pg/mL) (Table 3).\u003c/p\u003e\n\u003cp\u003eMoreover, PAL was considered an effect modifier of the association between serum 25(OH)D levels and IL-6 (p =0.03), IL-10 (p =0.008), IL-12p70 (p =0.014) and TNF-\u0026alpha; (p = 0.018) levels. Therefore, association analyses were performed separately in IA and PA (Fig. 3). In participants with a PAL greater than 150 minutes, IL-6, IL-10, TNF-\u0026alpha; and IL-12p70 levels tended to be lower as plasma 25(OH)D concentration increased (Fig. 4).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e: Coefficients of linear regression models for the association between 25(OH)D status and inflammatory markers in the total population and according to physical activity level. Health and Food Study (ESA - Vi\u0026ccedil;osa), 2012-2014\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"1040\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eInflammatory markers\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003eIL-6 (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003eIL-10 (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003eTNF-\u0026alpha; (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003eIL-8 (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003eIL-1\u0026beta; (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003eIL-12p70 (pg/mL)#\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003eus-CRP (mg/L) #\u003c/p\u003e\n \u003cp\u003e\u0026beta; (CI95%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e25(OH)D#\u003c/p\u003e\n \u003cp\u003eModel 1\u003c/p\u003e\n \u003cp\u003eModel 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.37 (-0.63; -0.10)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.29 (-0.52; -0.06)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.21(-0.37;-0.05)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.24(-0.41;-0.07)\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.29 (-0.49; -0.07)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.30 (-0.53; -0.07)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e-0.06 (-0.21; 0.09)\u003c/p\u003e\n \u003cp\u003e-0.01 (-0.17; 0.15)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.24 (-0.45;-0.04)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.25 (-0.47;-0.02)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.22 (-0.42; -0.02)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.22 (-0.44; -0.007)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.46 (-0.79;-0.12)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e-0.12 (-0.47; 0.22)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eInteraction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e25(OH)D*\u003c/p\u003e\n \u003cp\u003ePAL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.44 (-0.72; -0.17)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.38(-0.58; -0.1)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.46 (-0.73; -0.18)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e-0.07(-0.26; 0.11)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.46(-0.73;-0.18)*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e-0.34(-0.59; -0.06)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.03(-0.38; 0.43)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eIL: interleukin; BMI: body mass index; PAL: physical activity level; TNF:\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003etumor necrosis factor.\u003c/p\u003e\n\u003cp\u003e# log transformed. Model 1: Adjusted for age group and sex. Model 2: Adjusted for age group, sex, season, PAL, smoking, BMI, schooling and PTH.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e* PAL variable and interaction term were not significant in model 2\u003cstrong\u003e. (p=0.321)\u003c/strong\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study aimed to investigate the associations between 25(OH)D concentrations and inflammatory biomarkers in a Brazilian sample, and the presence of an interaction with physical activity and its mediating effect. We observed a negative association between 25(OH)D status and IL-6, IL-10, IL-1β, IL-12p70, and TNF-α in both models and us-CRP in the unadjusted model. This negative association remains in the presence of an interaction factor with PAL, confirming our hypothesis that higher 25(OH)D concentrations are associated with lower levels of inflammatory biomarkers in physically active adults.\u003c/p\u003e \u003cp\u003eElevated levels of inflammatory markers are associated with the pathogenesis of various metabolic disorders and non-communicable diseases. For instance, TNF-α and IL-1β play a role in the development of insulin resistance[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]; IL-6 has been linked to risk factors for cardiovascular diseases[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]; IL-12p70 has been identified as a critical marker in the early stages of atherosclerosis[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]; and CRP levels above 10 mg/L indicate a 4% increased risk of developing cardiovascular diseases[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. This underscores the importance of understanding the factors that modulate inflammation in order to prevent and manage inflammation-related disorders and improve overall health outcomes.\u003c/p\u003e \u003cp\u003eIn general, vitamin D is known for its multiple roles in the body, including regulation of the immune system and modulation of inflammation. The multiple roles of vitamin D in the immune system are well studied, especially the immunomodulatory effect on decreasing proinflammatory cytokine production, immunoglobulin release, and increasing anti-inflammatory cytokine secretion[\u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In our study, us-CRP was inversely associated with serum 25(OH)D status, and for each unit increase in 25(OH)D concentration, an average decrease in the inflammatory biomarkers IL-6, IL-10, TNF-α, IL-1β, and IL-12p70 was observed. The mechanism by which 25(OH)D affects inflammatory markers is still unclear. However, there is evidence that 25(OH)D can modulate the production of inflammatory cytokines both in vivo and in vitro, decreasing the levels of inflammatory cytokines and increasing the levels of anti-inflammatory markers [\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. In addition, a systematic review found that 25(OH)D reduces the production of pro-inflammatory cytokines, including IL-6 and TNF-α[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThere is some evidence to suggest that vitamin D deficiency may be associated with higher levels of inflammatory markers such as CRP and pro-inflammatory cytokines. However, the high prevalence of hypovitaminosis D observed reinforces that it is a global issue. The effectiveness of endogenous vitamin D (VD) production depends on environmental factors related to latitude, season and time of day of sun exposure, topical sunscreen use and skin pigmentation[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. In the present sample, even though the city of Vi\u0026ccedil;osa has a tropical climate, with mild winters and summers with abundant sunlight, a high prevalence of insufficient (44.7%) and deficient (11.6%) 25(OH)D levels was observed, with the lowest average observed in winter (25.49; 95%CI 23.89), compared to the other seasons. This is probably because in this season there is a lower incidence of UVB radiation and an increase in the use of clothing, consequently reducing the endogenous synthesis of vitamin D[\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHigher PAL have been associated with adequate 25(OH)D status through several plausible mechanisms. First, individuals who engage in regular exercise, particularly outdoor activities, tend to have greater sun exposure, thereby increasing cutaneous 25(OH)D synthesis under adequate ultraviolet B (UVB) radiation[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Second, exercise promotes lipolysis in adipose tissue, potentially mobilizing vitamin D stored in fat and increasing circulating 25(OH)D concentrations[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Additionally, physically active individuals often maintain healthier body composition and metabolic profiles, which may optimize vitamin D metabolism and utilization[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Finally, an active lifestyle is generally associated with other healthy habits, such as a balanced diet and lower rates of obesity, which in turn contribute to improved vitamin D status[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn our study, being physically active (PA) was associated with lower levels of IL-6. This inflammatory marker can act as both pro- and anti-inflammatory. Its ambiguous role depends on the stimulus that triggers its secretion, being an anti-inflammatory marker when secreted by skeletal muscle[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. A physically active lifestyle has been shown to lead to beneficial anti-inflammatory modulation. In general, the most sedentary group had higher odds of an elevated inflammatory status compared to the most active group[\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Thus, it appears that physical activity may help to increase 25(OH)D levels and reduce inflammation, thereby promoting an anti-inflammatory state. Panagiotakos et al.[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] and Scragg et al.[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] have shown that high levels of physical activity are associated with attenuated circulating levels of inflammatory cytokines and that highly active individuals have significantly increased circulating levels of 25(OH)D compared to inactive individuals, which is consistent with our findings.\u003c/p\u003e \u003cp\u003eHowever, there is controversy in the literature regarding the modulating effect of vitamin D, inflammatory markers and their mediation with physical activity levels, possibly due to methodological issues. Carrillo et al. (2012)[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] observed that 12 weeks of resistance training with (n\u0026thinsp;=\u0026thinsp;13, 53.8% women) or without (n\u0026thinsp;=\u0026thinsp;10, 50.0% women) vitamin D supplementation (4000 IU/day) did not improve IL-6, CRP or TNF-α. However, the sample was recruited in winter and the authors provided and instructed the sample to use sunscreen every day, which could have interfered with the effect of outdoor physical activity on 25(OH)D levels. Moreover, in addition to the resistance training intervention, the sample (BMI\u0026thinsp;\u0026gt;\u0026thinsp;25 kg/m\u003csup\u003e2\u003c/sup\u003e) was weight stable, which could mean that the mobilization of vitamin D stored in the body was impaired. In contrast, Nazarabadi et al. (2022)[\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] observed that in a sample of 46 postmenopausal women with metabolic syndrome, the intervention of physical activity or vitamin D supplementation (50000 IU/day) for 8 weeks decreased CRP and IL-6 levels and improved metabolic syndrome parameters. And the authors observed that the physical activity plus vitamin D supplementation group produced better results than physical activity or vitamin D supplementation alone.\u003c/p\u003e \u003cp\u003eTo understand the modulating effect of vitamin D, inflammatory markers and their mediation with the level of physical activity, an interaction term was included in the adjusted model in our study. As a result, we observed that the mean levels of inflammatory markers decreased in individuals who had higher levels of PAL and 25(OH)D concentration. Therefore, it seems that while vitamin D deficiency is associated with greater inflammation, physical activity tends to have a beneficial effect on both, potentially helping to increase vitamin D levels and reduce inflammatory markers.\u003c/p\u003e \u003cp\u003eThe present study had some limitations, such as its cross-sectional design, which did not allow inferences about the direction of the associations found. However, the cluster sampling and the population-based design mitigated this limitation. Although the IPAQ is widely used and standardized, it relies on self-reported information, which is subject to recall bias, over- or underestimation of activity duration and intensity, and social desirability bias. Furthermore, the IPAQ does not capture detailed temporal patterns of physical activity or account for activities of short duration but high intensity. For future studies aiming to obtain more objective and precise measures, direct assessment methods or other wearable devices are recommended. These devices can provide real-time data on activity intensity, frequency, and duration, thereby reducing measurement error and allowing for more accurate classification of PAL. Strengths of this study include the population-based sample size of 712 individuals and the methodological rigor applied in the data collection process, such as training the entire team involved and conducting a pilot study. These factors reinforced the quality and ensured the validity of the results obtained.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003e25(OH)D and physical activity play complementary roles in reducing inflammation. Controlling vitamin D deficiency and promoting an active lifestyle, preferably outdoors, may be critical strategies to improve population health by reducing the impact of elevated levels of inflammatory markers, potentially contributing to overall well-being.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research project was approved by the Research Ethics Committee of the Federal University of Viçosa (Official Letter 02/2013).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll participants signed the informed consent form.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone declared.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received financial support from the National Council for Scientific and Technological Development (CNPq process 481418/2011 3), the Research Support \u0026nbsp;Foundation of the State of Minas Gerais (FAPEMIG process APQ-00296 12).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGiana Zarbato Longo: Conceived the study idea, was responsible for statistical analysis and coordinator of the study.\u003c/p\u003e\n\u003cp\u003eDanielle Cristina Guimarães da Silva: data acquisition and wrote the main text.\u003c/p\u003e\n\u003cp\u003eMariana Papinni Gabiatti: made many useful suggestions during the revision of the\u003c/p\u003e\n\u003cp\u003emanuscript.\u003c/p\u003e\n\u003cp\u003eFernanda Hansen: made critical comments and revisions for the initial manuscript.\u003c/p\u003e\n\u003cp\u003eFabrícia Geralda Ferreira: wrote the initial manuscript.\u003c/p\u003e\n\u003cp\u003eDiego Augusto Santos Silva: physical education expertise and significant reviewer.\u003c/p\u003e\n\u003cp\u003eLeandro Licursi de Oliveira: performed the biochemical analysis of cytokines.\u003c/p\u003e\n\u003cp\u003eAmanda Alcaraz da Silva: wrote the discussion text.\u003c/p\u003e\n\u003cp\u003eYara Maria Franco Moreno: made critical comments and revisions for the initial manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to thank all of the participants for their dedication National Council for Scientific and Technological Development and also the Foundations for Supporting Research in the states of Minas Gerais for financially funding the project. The federal university of viçosa and its facilities for the study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eHolick MF, Chen TC. Vitamin D deficiency: A worldwide problem with health consequences. Am J Clin Nutr 2008;87:1080\u0026ndash;6. https://doi.org/10.1093/ajcn/87.4.1080s.\u003c/li\u003e\n \u003cli\u003ePereira-Santos M, Santos JYG dos, Carvalho GQ, Santos DB dos, Oliveira AM. Epidemiology of vitamin D insufficiency and deficiency in a population in a sunny country: Geospatial meta-analysis in Brazil. Crit Rev Food Sci Nutr 2019;59:2102\u0026ndash;9. https://doi.org/10.1080/10408398.2018.1437711.\u003c/li\u003e\n \u003cli\u003ede Sire A, Gallelli L, Marotta N, Lippi L, Fusco N, Calafiore D, et al. Vitamin D Deficiency in Women with Breast Cancer: A Correlation with Osteoporosis? A Machine Learning Approach with Multiple Factor Analysis. 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World Heal Organ 2020:535.\u003c/li\u003e\n \u003cli\u003eMatsudo S, Ara\u0026uacute;jo T, Matsudo V, Andrade D, Andrade E, Oliveira LC, et al. Question\u0026aacute;rio Internacional De Atividade F\u0026iacute;sica (Ipaq): Estupo De Validade E Reprodutibilidade No Brasil. Rev Bras Atividade F\u0026iacute;sica Sa\u0026uacute;de 2012;6:5\u0026ndash;18. https://doi.org/10.12820/rbafs.v.6n2p5-18.\u003c/li\u003e\n \u003cli\u003ePludowski P, Takacs I, Boyanov M, Belaya Z, Diaconu CC, Mokhort T, et al. Clinical Practice in the Prevention, Diagnosis and Treatment of Vitamin D Deficiency: A Central and Eastern European Expert Consensus Statement. Nutrients 2022;14:1\u0026ndash;18. https://doi.org/10.3390/nu14071483.\u003c/li\u003e\n \u003cli\u003eHolick MF. Resurrection of vitamin D deficiency and rickets. 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The impact of C reactive protein on global cardiovascular risk on patients with coronary artery disease. Curr Heal Sci J 2013;39:225\u0026ndash;31.\u003c/li\u003e\n \u003cli\u003eLemire JM. lmmunomodulatory Role of 1 , 25-Dihydroxyvitamin D3 1984;31:26\u0026ndash;31.\u003c/li\u003e\n \u003cli\u003eZhang Y, Leung DYM, Richers BN, Liu Y, Remigio LK, Riches DW, et al. Vitamin D Inhibits Monocyte/Macrophage Proinflammatory Cytokine Production by Targeting MAPK Phosphatase-1. J Immunol 2012;188:2127\u0026ndash;35. https://doi.org/10.4049/jimmunol.1102412.\u003c/li\u003e\n \u003cli\u003eChen S, Lipsky PE, Chen S, Sims GP, Chen XX, Gu YY. on Human B Cell Differentiation 3 D Modulatory Effects of 1,25-Dihydroxyvitamin Modulatory Effects of 1,25-Dihydroxyvitamin D 3 on Human B Cell Differentiation. J Immunol Ref J Immunol Samsung Med Cent 2007;179:1634\u0026ndash;47.\u003c/li\u003e\n \u003cli\u003eAkbari M, Ostadmohammadi V, Lankarani KB, Tabrizi R, Kolahdooz F, Heydari ST, et al. The Effects of Vitamin D Supplementation on Biomarkers of Inflammation and Oxidative Stress among Women with Polycystic Ovary Syndrome: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. Horm Metab Res 2018;50:271\u0026ndash;9. https://doi.org/10.1055/s-0044-101355.\u003c/li\u003e\n \u003cli\u003eMansournia MA, Ostadmohammadi V, Doosti-Irani A, Ghayour-Mobarhan M, Ferns G, Akbari H, et al. Abbreviations GSH Total glutathione hs-CRP High-sensitivity C-reactive protein MDA Malondialdehyde NO Nitric oxide TAC Total antioxidant capacity The Effects of Vitamin D Supplementation on Biomarkers of Inflammation and Oxidative Stress in Diabetic Patie. Vitam D Diabetes \u0026hellip; Horm Metab Res Horm Metab Res Mansournia MA Al Vitam D Diabetes \u0026hellip; Horm Metab Res 2018;50:429\u0026ndash;40.\u003c/li\u003e\n \u003cli\u003eSepidarkish M, Farsi F, Akbari-Fakhrabadi M, Namazi N, Almasi-Hashiani A, Maleki Hagiagha A, et al. 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Am J Epidemiol 2008;168:577\u0026ndash;86. https://doi.org/10.1093/aje/kwn163.\u003c/li\u003e\n \u003cli\u003eSegheto KJ, Pereira M, Da Silva DCG, De Carvalho CJ, Massardi FR, Kakehasi AM, et al. Vitamin d and bone health in adults: A systematic review and meta-analysis. Cienc e Saude Coletiva 2021;26:3221\u0026ndash;44. https://doi.org/10.1590/1413-81232021268.15012020.\u003c/li\u003e\n \u003cli\u003eVillar-Fincheira P, Sanhueza-Olivares F, Norambuena-Soto I, Cancino-Arenas N, Hernandez-Vargas F, Troncoso R, et al. Role of Interleukin-6 in Vascular Health and Disease. Front Mol Biosci 2021;8:1\u0026ndash;11. https://doi.org/10.3389/fmolb.2021.641734.\u003c/li\u003e\n \u003cli\u003eCho SMJ, Lee H, Shim JS, Jeon JY KH. Association between physical activity and inflammatory markers in community-dwelling, middle-aged adults. Appl Physiol Nutr Metab 2021;33566730:1\u0026ndash;31. https://doi.org/10.1139/apnm-2020-1069.\u003c/li\u003e\n \u003cli\u003ePanagiotakos DB, Pitsavos C, Chrysohoou C, Kavouras S, Stefanadis C. The associations between leisure-time physical activity and inflammatory and coagulation markers related to cardiovascular disease: The ATTICA Study. Prev Med (Baltim) 2005;40:432\u0026ndash;7. https://doi.org/10.1016/j.ypmed.2004.07.010.\u003c/li\u003e\n \u003cli\u003eNazarabadi PN, Etemad Z, Hoseini R MF. Anti-Inflammatory Effects of a Period of Aerobic Training and Vitamin D Supplementation in Postmenopausal Women with Metabolic Syndrome. Int J Prev Med 2022;13:1\u0026ndash;7.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"25-hydroxyvitamin D, cytokines, inflammation, physical activity, nutritional epidemiology.","lastPublishedDoi":"10.21203/rs.3.rs-6214919/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6214919/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eVitamin D is of vital importance for bone health and also regulates inflammatory cytokines, which contribute to immune signaling and defenses. It is possible that physical activity could influence serum 25(OH)D levels, and it has been suggested that exercise can increase serum 25(OH)D concentration. The aim of this study was to investigate the association between inflammatory biomarkers and 25(OH)D concentration in a sample of Brazilian adults, taking into account the influence of physical activity on this association.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eThis is a cross-sectional population-based study, conducted with 712 adults aged 20 to 59 years living in the urban area of Viçosa, Minas Gerais, Brazil. Sociodemographic, anthropometric, behavioral and biochemical variables were collected. Multiple linear regression models (crude and adjusted) were used to determine associations between 25(OH)D concentration and inflammatory biomarkers, with statistical significance analyzed at 5%.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003cem\u003e \u003c/em\u003eIn the multiple linear regression models, it can be observed that for each unit increase in the concentration of 25(OH)D, there was an average decrease in the inflammatory biomarkers. Physical activity level was considered an effect modifier of the association between serum 25(OH)D levels and some cytokines, and participants whose PAL was greater than 150 minutes tended to have lower levels of IL-6, IL-10, TNF-α and IL-12p70 when plasma 25(OH)D concentration was increased.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e\u003cem\u003e \u003c/em\u003eHigher 25(OH)D concentrations were associated with lower levels of inflammatory biomarkers in Brazilian adults. Furthermore, physical activity served as a significant effect modifier of this association, enhancing the reduction of pro-inflammatory cytokines. These results underscore the importance of maintaining adequate 25(OH)D levels in conjunction with an active lifestyle to modulate inflammation and promote overall health.\u003c/p\u003e","manuscriptTitle":"Association between Cytokines, 25-Hydroxyvitamin D, and Physical Activity: Evidence from a cross sectional Brazilian Population","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-20 09:24:54","doi":"10.21203/rs.3.rs-6214919/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":"b33224ec-62c5-4560-9a9e-15f1b825b2a8","owner":[],"postedDate":"March 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-03-24T04:38:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-20 09:24:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6214919","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6214919","identity":"rs-6214919","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}