Expression of myostatin and follistatin genes in hypothyroid rats following a moderate-intensity exercise period

Shamsi, Behnam; Saremi, Abbas; Malekipooya, Mohammad

doi:10.22034/27.2.25

[Home ] [Archive]

[ فارسی ]

Main Menu

Home

Journal Information

Articles archive

For Authors

For Reviewers

Ethics

Contact us

Search in website

Receive site information

Index

‌

Volume 27, Issue 2 (7-2025)

EBNESINA 2025, 27(2): 25-36

Back to browse issues page

Expression of myostatin and follistatin genes in hypothyroid rats following a moderate-intensity exercise period

Behnam Shamsi

, Abbas Saremi ^*

, Mohammad Malekipooya

Department of Exercise Physiology, Faculty of Sports Sciences, Arak University, Arak, Iran , a-saremi@araku.ac.ir

Keywords: Hypothyroidism, Exercise, Myostatin, Follistatin

Full-Text [PDF 1746 kb] (66 Downloads) | Abstract (HTML) (208 Views)

Type of Study: Original | Subject: Military Medicine
Received: 2024/11/30 | Revised: 2025/09/29 | Accepted: 2025/04/19 | Published: 2025/06/22

Extended Abstract: (30 Views)

Introduction

Hypothyroidism (HO) stands as one of the most prevalent endocrine disorders, characterized by diminished thyroid hormone levels. The etiology of this condition is multifaceted, with contributing factors including autoimmune thyroiditis, iodine deficiency, surgical interventions, or radioiodine therapy [1]. A mild form of hypothyroidism is identified when thyroxine (T4) levels remain within the normal range, while thyroid-stimulating hormone (TSH) levels are elevated but remain below 10 mIU/L [3]. The interplay between thyroid hormones, TSH levels, and muscle structure is well-documented, underscoring the critical role of thyroid hormones (THs) as regulators of muscle metabolism in both health and disease. These hormones function as potent anabolic and catabolic agents, making their investigation pivotal in understanding muscle function [5].
Previous research has explored the effects of exercise training on myostatin and follistatin levels, revealing no significant alterations in these markers across various training sessions [17]. Given this context, the present study sought to elucidate the changes in myostatin and follistatin gene expression following a regimen of moderate-intensity exercise (MIT) in rats with induced hypothyroidism. This investigation aimed to address a potential gap in the literature by examining how exercise influences these molecular markers in a hypothyroid state.

Methods

The study employed 21 male Wistar rats, aged eight weeks, with an average weight of 210 ± 15 grams. After acclimatization to the laboratory environment, the animals were randomly allocated into three groups of seven: a healthy control group (H), a hypothyroid control group (HO), and a hypothyroid group subjected to endurance exercise (HO+EX). Hypothyroidism was induced via intraperitoneal injection of methimazole powder over a 10-day period [18]. The endurance exercise protocol was conducted over six weeks, with sessions occurring five days per week [22].
Two days after the completion of the exercise protocol, the rats were anesthetized and euthanized using an intraperitoneal injection of ketamine (75 mg/kg) and xylazine (10 mg/kg) to facilitate the extraction of muscle tissue. RNA and cDNA were subsequently isolated from the muscle samples, and quantitative gene expression analysis was performed using PCR with the 2^(-ΔΔCt) method. The Shapiro-Wilk test was employed to assess the normal distribution of the data. Statistical comparisons between groups were conducted using one-way analysis of variance (ANOVA), followed by Tukey's post hoc test for pairwise comparisons.

Results

The findings revealed that the induction of hypothyroidism resulted in a significant increase in myostatin levels (p < 0.0001) and a non-significant decrease in follistatin levels (p = 0.353) among the rats. Conversely, endurance training led to a marked reduction in myostatin levels (p < 0.0001) and a significant elevation in follistatin levels (p = 0.01) in the hypothyroid rats. These results suggest that moderate-intensity exercise exerts a regulatory effect on the expression of these genes in the context of hypothyroidism.

Discussion and Conclusion

The key outcomes of this study include a substantial reduction in myostatin gene expression and a notable increase in follistatin gene expression compared to the hypothyroid control group. These findings align with prior research by Carneiro et al., who identified myostatin as a critical regulator of muscle growth. Their work demonstrated that hypothyroid patients exhibit elevated myostatin expression, which correlates with muscle atrophy [10]. This underscores the therapeutic potential of physical activity in mitigating muscle deterioration in hypothyroid individuals.
Further supporting these observations, Rashidlamir et al. investigated the impact of endurance training on myostatin protein expression in cardiac muscle and reported a significant decline in myostatin levels post-exercise [24]. Similarly, Asad et al. observed a reduction in myostatin concentration following a combined exercise program in non-athletic men [26]. Mechanistically, myostatin is known to inhibit skeletal muscle growth by activating the MSTN/SMAD signaling pathway [28].
The study also highlighted the role of follistatin, which exhibited a significant increase in expression following endurance training. This finding is consistent with the work of Karimi et al., who documented elevated follistatin gene expression in response to exercise [34]. Additionally, Elliott et al. reported a rise in follistatin levels among non-athletic elderly participants after high-intensity exercise training [35]. Hansen et al. further demonstrated that plasma follistatin secretion increases during physical activities such as cycling, knee bending, and swimming [36]. Follistatin appears to function by inhibiting activin and attenuating myostatin signaling, positioning it as a promising therapeutic target for muscle growth regulation, metabolic syndrome, and inflammation.
In conclusion, the study demonstrates that moderate-intensity endurance training induces favorable changes in myostatin and follistatin gene expression in hypothyroid rats. These molecular adaptations suggest that exercise may serve as a viable intervention to counteract muscle atrophy and wasting in hypothyroid individuals.

Ethical Considerations

This study received approval from the Ethics Committee for Biomedical Research at Islamic Azad University, Arak Branch, under the ethics code IR.IAU.ARAK.REC.1403.273. All procedures adhered to ethical guidelines to ensure the humane treatment of the animals involved.

Funding

There is no funding support.

Authors’ Contribution

Authors contributed equally to the conceptualization and writing of the article. All of the authors approved the content of the manuscript and agreed on all aspects of the work.

Conflict of Interest

Authors declared no conflict of interest.

Acknowledgments

The authors would like to express their gratitude to all those who contributed to this research project. This article is the result of the first author's postdoctoral research project.

References

1. 1. Taylor PN AD, Scholz A, Gutierrez- Buey G, Lazarus JH, Dayan CM, et al. . Global epidemiology of hyperthyroidism and hypo-thyroidism. Nat Rev Endocrinol. 2018;14:30116.

2. 2. B. G. Review. Why can insulin resistancebe a natural consequence of thyroid dysfunction? thyroid Res 2011; 9(152850).

3. 3. M. S. The thyroid hormone breakthrough; overcoming sexual and hormonal problems at every age. . Tehran Katibe Mehr. 2006;256.

4. 4. Kalantar Hormozi M NPE, Asadi M, Asadi Pooya K, Zende Boody S, Ranjbar EmraniGh. . Sub clinical hypothyroidism. . Iranian South Medical journal. 2011;14:51-60 (Persian).

5. 5. Rooyackers OE NK. Hormonal regulation of human muscle protein metabolism. Annu Rev Nutr. 1997;17: 457-85.

6. 6. Ma K MC, Bhasin S, et al. Glucocorticoid-induced skeletal muscle atrophy is associated with upregulation of myostatin gene expression. . Am J Physiol Endocrinol Metab 2003;285:E363-71.

7. 7. Saunders MA, Good JM, Lawrence EC, Ferrell RE, Li WH, Nachman MW. Human adaptive evolution at Myostatin (GDF8), a regulator of muscle growth. Am J Hum Genet. 2006;79(6):1089-97.

8. 8. Carneiro IC-P, A. Muñoz, J.L. Labandeira-García, J. Devesa, and V.M. Arce. . Hypothyroidism is associated with increased myostatin expression in rats. . J Endocrinol Invest. 2008;31:773-8.

9. 9. Lagally KM, & Robertson, R. J. (). . Construct validity of the OMNI resistance exercise scale. . Journal of Strength and Conditioning Research. 2006;20(2):252

10. 10. Willoughby DS. Effects of an alleged myostatin-binding supplement and heavy resistance training on serum myostatin, muscle strength and mass, and body composition. . International ournal of Sport Nutrition & Exercise Metabolism. 2004 14(4).

11. 11. Tortoriello DV, Sidis Y, Holtzman DA, Holmes WE, Schneyer AL. Human follistatin-related protein: a structural homologue of follistatin with nuclear localization. Endocrinology. 2001;142(8):3426-34.

12. 12. Rodino-Klapac LR, Haidet AM, Kota J, Handy C, Kaspar BK, Mendell JR. Inhibition of myostatin with emphasis on follistatin as a therapy for muscle disease. Muscle Nerve. 2009;39(3):283-96.

13. 13. Assad MaJ. The effect of combined exercise program (resistance+ endurance) on plasma myostatin levels in non-athlete obese men. . Sports Life Scienc. 2012;15:77-89.

14. 14. Esazadeh L HKA, Khajeie R & Hejazi S.M. . The effect of combined exercise sequence (resistance-aerobic) on some factors of physical fitness, functional capacity and serum levels of myostatin and follicatin hormones in postmenopausal women (clinical trial). . Journal of Sports Life Sciences. 2020;12 (2):189-206. .

15. 15. Bagheri ea. The effect of combined training sequence (strength and endurance) on serum levels of myostatin, folistatin and ratio of folistatin on myostatin in elderly women. . Sports Physiology 2015;7(26):143-64.

16. 16. Siqueira CC RR, Tiengo AN, Tufik S, Schenberg LC. . Methimazole-induced hypothyroidism inhibits the panic-like behaviors produced by electrical stimulation of dorsal periaqueductal gray matter of rats. . Psychoneuroendocrinology 2010;35(3):706-16.

17. 17. DS. C. Antithyroid drugs. . Rev N EngL J Med 2005;352:905-17.

18. 18. M. M. The response of fibroblast-derived factor and endostatin to acute rehabilitation of physical activity along with electrical stimulation in infarcted rats. . Medical Journal of Mashhad University of Medical Science. 2025;67 (3).

19. 19. M. M. Response of acute incremental aerobic activity along with electrical stimulation on some markers of angiogenesis in Isoproterenol induced rats. . EBNESINA. 2024;26 (1):28-37.

20. 20. Schefer V TM. Oxygen consumption in adult and AGED C57BL/6J mice during acute treadmill exercise of different intensity. Exp Gerontol. 1996;31(3):387-92.

21. 21. Fathi M MZM, Khairabadi S, Hejazi K. Effect of Aerobic Exercise on Thyroid Hormones and Quality of Life in Obese Postmenopausal Women.. . mljgoums. 2018;12(6):5-11.

22. 22. Rashidlamir A HS, Hejazi K, Anberani SM. . The effect of eight weeks resistance and aerobic training on myostatin and follistatin expression in cardiac muscle of rats. . Journal of Cardiovascular and Thoracic Research. 2016;8(4):164.

23. 23. Jokar M AAM, Sharafati M.M. . The effect of endurance training on the content of myostatin and SMAD2/3 proteins in the left ventricular tissue of heart muscle of type 1 diabetic rats. Iranian Journal of Diabetes and Metabolism (Iranian Journal of Diabetes and Lipid 2019;3(4):191-9.

24. 24. Asad MaJV. The Effect of Concurrent (Resistance and Endurance) Training on Plasma Myostatin Levels in Obese Non-Athlete Men. . Journal of Sport Biosciences, . 2013;4(15):77-89.

25. 25. Hittel DS AM, Sarna N, Shearer J, Huffman KM, Kraus WE. . Myostatin decreases with aerobic exercise and associates with insulin resistance. . Medicine and Science in Sports And Exercise 2010;42(11):2023.

26. 26. Fedoruk MaJR. Myostatin inhibition: a potential performance enhancement strategy? . Scandinavian journal of medicine & science in sports 2008;18(2):123-31.

27. 27. McFarlane C, et al., . Myostatin signals through Pax7 to regulate satellite cell self-renewal. . Experimental cell research. 2008;314(2):317-29.

28. 28. Wullschleger S, R. Loewith, and M.N. Hall, . TOR signaling in growth and metabolism. . Cell. 2006;124(3):471-84.

29. 29. Carlson CJ, F.W. Booth, and S.E. Gordon, . Skeletal muscle myostatin mRNA expression is fiber-type specific and increases during hindlimb unloading. . American Journal of Physiology Regulatory, Integrative and Comparative Physiology. 1999 277(2):R601-R6. .

30. 30. Latres E, et al., . Myostatin blockade with a fully human monoclonal antibody induces muscle hypertrophy and reverses muscle atrophy in young and aged mice. . Skeletal Muscle,. 2015;5(1):1-13.

31. 31. Bahram M E HS, vahedi cole sara N, Afroundeh R, davarpanah S. . The effect of training TRX on serum levels of myostatin and Follistatin and neuromuscular function in overweight elderly men. IJRN 2023;9(3):88-98.

32. 32. Karimi R, Fakhrpour, R., Zarneshan, A. . Effect of Resistance Training with Milk Protein Concentrate (MPC) Supplementation on serum levels of Follistatin and myostatin and Muscle Hypertrophy in Beginner Bodybuilders. . Journal of Applied Health Studies in Sport Physiology. 2022;9(1):151-63.

33. 33. Elliott BT HP, Sculthorpe N, Grace FM, Stratton D, Hayes LD. . Lifelong exercise, but not short‐ term high‐ intensity interval training, increases GDF11, a marker of successful aging: a preliminary investigation. . Physiological Reports. 2017;5(13):e13343.

34. 34. Hansen J BC, Nielsen AR, Hojman P, Whitham M, Febbraio MA, et al. . Exercise induces a marked increase in plasma follistatin: evidence that follistatin is a contraction-induced hepatokine. Endocrinology. 2011;152(1):164-71.

35. 35. Biglari S GAA, Kordi M.R, Ghardashi Afousi A. . The Effect of 8 Weeks High-intensity Interval Training on Myostatin and Follistatin Gene Expression in Gastrocnemius Muscle of the Rats. . Arak Medical University Journal (AMUJ). 2018;21(130):1-10. .

36. 36. Anastasilakis AD, et al., . Circulating follistatin displays a day-night rhythm and is associated with muscle mass and circulating leptin levels in healthy, young humans. Metabolism. 2016;65(10):1459-65.

37. 37. Tao RW, Caixia; Stöhr, Oliver; Qiu, Wei; Hu, Yue; Miao, Ji; Dong, X. Charlie; Leng, Sining; Stefater, Margaret; Stylopoulos, Nicholas; Lin, Lin; Copps, Kyle D.; White, Morris F. . Inactivating hepatic follistatin alleviates hyperglycemia. . Nature Medicine. 2018;24(7):1058-69.

Send email to the article author

Add your comments about this article

‎ 10.22034/27.2.25

Ethics code: IR.IAU.ARAK.REC.1403.273

Mendeley

Zotero

RefWorks

Shamsi B, Saremi A, Malekipooya M. Expression of myostatin and follistatin genes in hypothyroid rats following a moderate-intensity exercise period. EBNESINA 2025; 27 (2) :25-36
URL: http://ebnesina.ajaums.ac.ir/article-1-1377-en.html

Rights and permissions
	This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Volume 27, Issue 2 (7-2025)

Back to browse issues page

Persian site map - English site map - Created in 0.07 seconds with 39 queries by YEKTAWEB 4722