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Combined Effects of High-Intensity Interval Training and Vitamin D Supplementation on the Levels of Inflammatory Markers, Transforming Growth Factor Beta 1 (TGF-β1) and Tumor Necrosis Factor Alpha (TNF-α), in Young Women with Vitamin D Deficiency: A Clinical Trial
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Marzieh Noshirvanpour Kochaksaraie1    , Hossain Arabzadeh2 , Masoumeh Habibian *3 |
1- M.Sc in Exercise Physiology, Department of Physical Education and Sport Sciences, QaS.C., Islamic Azad University, Qaemshahr, Iran.
2- M.Sc in Exercise Biomechanic, Department of Motor Behavior and Sports Biomechanics, Faculty of Physical Education and Sport Sciences, University of Mazandaran, Mazandaran, Iran.
3- Associate Professor, Department of Physical Education and Sport Sciences, QaS.C., Islamic Azad University, Qaemshahr, Iran. , habibian_m@yahoo.com |
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Keywords: High-Intensity Interval Training [MeSH], Tumor Necrosis Factor-alpha [MeSH], Transforming Growth Factor beta1 [MeSH], Vitamin D Deficiency [MeSH]
Article ID: Vol27-22 |
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Type of Study: Original Articles |
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Exercise Physiology
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Abstract: (78 Views) |
Extended Abstract
Introduction
Obesity is recognized as a chronic inflammatory disease, with transforming growth factor beta 1 (TGF-β1) playing a significant role in its pathogenesis, as well as in the regulation of adipocyte differentiation and the immuno-inflammatory microenvironment. Tumor necrosis factor alpha (TNF-α) and TGF-β are among the main adipokines involved in regulating inflammatory processes in obese individuals. TGF-β1 is a polypeptide member of the cytokine family. The TGF-β superfamily comprises a large group of critical cellular proteins that regulate a wide range of cellular processes by signaling through high-affinity TGF-β receptors. TGF-β1 is fundamentally expressed in adipose tissue, cultured preadipocytes, and differentiated adipocytes. TGF-β1 signaling regulates many cellular processes, such as autophagy, apoptosis, inflammation, fibrosis, and adipocyte differentiation.
TNF-α is a classic pleiotropic proinflammatory cytokine produced by adipose tissue. This cytokine is upregulated in obesity and contributes to obesity-related metabolic disease. Additionally, TNF-α can lead to hyperlipidemia by stimulating hepatic lipogenesis, promoting fat lipolysis alongside the suppression of triglyceride clearance, and inhibiting insulin-stimulated lipogenesis in adipose tissue. TNF-α directly alters the lipid metabolism of adipocytes by inhibiting the uptake of free fatty acids (FFAs) and lipogenesis, as well as by stimulating the release of FFAs via lipolysis. Higher levels of TNF-α have been observed in overweight and obese individuals compared to normal-weight individuals, indicating the cytokine's role in metabolic disorders and immune system activation within the adipose tissue.
Inadequate levels and deficiency of vitamin D represent a global health concern, with obese individuals being particularly vulnerable. This increased obesity-related vitamin D deficiency has been observed across various populations and age groups. Low levels of 25-hydroxyvitamin D are associated with higher levels of systemic inflammation. Evidence suggests that vitamin D can reduce inflammatory cytokines and consequently decrease systemic inflammation through binding to vitamin D receptors. One of the mechanisms for vitamin D deficiency and insufficiency in obesity is the sequestration of vitamin D in adipose tissue, which in turn culminates in vitamin D deficiency and subsequent obesity-induced inflammation. Furthermore, vitamin D deficiency exacerbates the activation of the TGF-β/Smad2/3 pathway. Two weeks of oral vitamin D3 supplementation has been shown to significantly reduce TNF-α serum levels in patients with intestinal inflammation.
Physical activity is considered a major factor in the treatment of obesity and its related diseases. For instance, the risk of morbidity and mortality is lower in physically active overweight or obese men and women compared to their sedentary counterparts. Furthermore, regular exercise training, in addition to reducing pro-inflammatory cytokines, leads to an increase in the circulating concentration of anti-inflammatory cytokines. Nevertheless, the effects of exercise training on the levels of the inflammatory markers TNF-α and TGF-β1 have been reported inconsistently.
This study was conducted to determine the combined effects of high-intensity interval training (HIIT) and vitamin D supplementation on the levels of the inflammatory markers TGF-β1 and TNF-α in young women with vitamin D deficiency.
Methods
This clinical trial was conducted on 39 sedentary, overweight young women with a body mass index (BMI) equal to or greater than 25 and less than 29 kg/m2 (age range= 23 to 29 years) during 2021. The inclusion criteria comprised sedentary, overweight women who had not participated in any regular physical activity for at least six months prior and whose 25-hydroxyvitamin D levels were less than 20 ng/mL. The exclusion criteria included cardiovascular diseases, hypertension, and inflammatory diseases, use of specific medications or supplements, pregnancy, failure to attend at least three regular training sessions, and non-adherence to the prescribed vitamin D supplementation regimen.
Participants were enrolled in the study following an introduction to the research procedures and completion of an informed consent form. They were asked to record every food item consumed over a 3-day period. Based on this record, the participants' mean daily caloric intake was calculated. Subsequently, an individual nutritional counseling session was conducted with each participant to ensure adherence to a consistent diet for weight maintenance. Participants were advised to continue their usual diet throughout the study period (particularly in the phases leading up to the blood sampling).
The subjects were selected using convenience and purposeful sampling and then randomly assigned, via a simple lottery method, into three groups of 13 participants each, as follows:
A control group: Placebo + food (once a week for 8 weeks)
Intervention group 1: HIIR-based running and placebo + food (once a week for 8 weeks)
Intervention group 2: HIIR-based running and vitamin D supplementation (a dose of 5,000 IU) with food (once a week for 8 weeks)
The placebo consisted of an oral paraffin pearl (manufactured by Zahravi Pharmaceutical Company, Iran), which was visually identical to the vitamin D supplementation.
Compliance with the consumption of the placebo and vitamin D supplementation by the subjects was reminded through weekly phone contacts.
A week prior to the start of the protocol, subjects were familiarized with how to perform the training. The structure of each training session for the two training intervention groups consisted of three parts: A warm-up (5 minutes), HIIT, and a cool-down (return to baseline) (5 minutes) as three sessions per week. The warm-up and cool-down were performed at an intensity of 50% of maximum heart rate (HRmax). The main training consisted of 12 one-minute repetitions of running at an intensity of 80% to 90% of HRmax, followed by one minute of active recovery at an intensity of 50% of HRmax.
The training sessions began in the first week with an intensity of 80% of HRmax and 6 repetitions. This gradually increased, reaching an intensity of 90% of HRmax with 12 repetitions by the fifth week (with a progressive increment of 5% in training intensity and 3 repetitions every two weeks) and continued until the final week. Furthermore, participants' HRmax was determined using the formula (220 minus age). Training intensity during the field-based exercises was monitored and controlled using Polar heart rate monitors or wristwatches.
Blood samples (5 mL) were collected from subjects, following 12 hours of overnight fasting (consumption of a light meal the night before blood sampling), in the two phases of pre-test and post-test (48 hours after the final training session). Samples were drawn in the morning from the antecubital vein of the left arm while the subject was in a seated position and after a 15-minute rest period. To separate the serum, blood samples were centrifuged for 15 minutes at 3000 rpm. The resulting serum was then transferred into microtubes, frozen at −80℃, and stored for the measurement of the research variables.
Pre-test 25-hydroxyvitamin D levels were determined using an enzyme-linked immunosorbent assay (ELISA) commercial kit (manufactured by Padtan Gostar Isar Company, Iran) with a sensitivity of 2.6 ng/mL. Serum concentrations of TGF-β1 and TNF-α were measured using ELISA commercial kits from China, with sensitivities of 0.75 ng/mL and 4.69 pg/mL, respectively.
Results
A statistically significant decrease was observed in the post-test TNF-α and TGF-β1 levels in the HIIT-based running group (5.10±0.62 and 17.73±1.63, respectively) and the HIIT-based running + vitamin D group (4.73±0.47 and 17.22±1.32, respectively) (P<0.001); however, the levels of these variables showed no statistically significant changes in the control group.
Pairwise comparisons of the mean percentage changes in TNF-α and TGF-β1 levels, conducted using the Mann-Whitney U test, demonstrated that intervention groups 1 and 2 differed significantly from the control group (P<0.001). Furthermore, the reduction in TNF-α and TGF-β1 levels was significantly greater in intervention group 2 (training + vitamin D) compared to intervention group 1 (training + placebo) (P<0.001).
Conclusion
Following 8 weeks of HIIT-based running, both inflammatory markers TGF-β1 and TNF-α decreased in the study subjects, and this reduction was more pronounced in the vitamin D-receiving group.
Regular physical exercise can inhibit the latent activation of TGF-β1. Besides the direct and reciprocal stimulation of TNF-α and TGF-β, TNF-α affects TGF-β receptors. TNF-α can increase the expression of TGF-β receptors type I and II and stimulate the phosphorylation of Smad3 in fibroblasts. In addition, TNF-α induces the expression of activator protein 1 (AP-1). This protein, upon binding to DNA, leads to increased transcription of the TGF-β1 gene. The TNF-α-induced increase in TGF-β1 expression may be associated with the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway. TGF-β1 and TNF-α have a reciprocal stimulatory effect, and their synergistic effects are primarily in enhancing the release of inflammatory factors. Therefore, it appears that HIIR-based running, by suppressing both of these inflammatory markers and inhibiting their synergistic effects, ultimately reduces inflammation in conditions of overweight combined with vitamin D deficiency.
The greater effect of combined intervention of HIIT-based running and vitamin D supplementation on the downregulation of TGF-β1 and TNF-α compared to HIIT-based running alone is among the other important findings of the present research.
Ethical Statement
This study received approval from the Ethics Committee of Islamic Azad University, Sari Branch (IR.IAU.SARI.REC.1402.309). It was also registered with the Iranian Registry of Clinical Trials (IRCT) (IRCT20190831044650N3).
Conflicts of Interest
No conflict of interest.
Acknowledgments
This article has been extracted from the master’s thesis of Ms. Marzieh Noshirvanpour Kochaksaraie in Exercise Physiology at the Faculty of Humanities, Islamic Azad University, Ghaemshahr Branch. We would like to thank all participants who sincerely assisted us in conducting this study.
Authors' Contributions
Marzieh Noshirvanpour Kochaksaraie (M.Sc): Project execution, Data collection, Drafting of the initial manuscript, Approval of the final manuscript.
Hossain Arabzadeh (M.Sc): Project execution, Data collection, Drafting of the initial manuscript, Approval of the final manuscript.
Masoumeh Habibian (Ph.D): Project administration and design, Data analysis, Interpretation of the results, Approval of the final manuscript.
Key Message: The combined intervention of HIIT-based running and vitamin D supplementation is more effective than HIIT-based running alone in strengthening the reduction of systemic inflammation.
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Noshirvanpour Kochaksaraie M, Arabzadeh H, Habibian M. Combined Effects of High-Intensity Interval Training and Vitamin D Supplementation on the Levels of Inflammatory Markers, Transforming Growth Factor Beta 1 (TGF-β1) and Tumor Necrosis Factor Alpha (TNF-α), in Young Women with Vitamin D Deficiency: A Clinical Trial. J Gorgan Univ Med Sci 2025; 27 (3) :10-20 URL: http://goums.ac.ir/journal/article-1-4561-en.html
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