The effect of regular resistance exercise, vitamin D, and calcium supplements on the gastrocnemius muscle in rats in the post-menopausal period: An experimental study


Background: Menopause is the natural termination of menstruation which affects the quality and important aspects of women’s life.

Objective: To evaluate the effect of regular resistance training (Ex) with vitamin D (Vit. D) and calcium (Ca) supplements in the postmenopausal period on muscle tissue in rats.

Materials and Methods: In this experimental study, 72 female Wistar rats (8-10-wk old) were randomly divided into control, placebo, Vit. D, Ca, Ex, Ca + Vit. D, Ex + Ca, Ex + Vit. D, and Ex + Ca + Vit. D groups. Control and placebo groups were fed with a standard diet and sesame oil, respectively. Two months after the ovariectomy, Ex, Ca (35 mg/kg), and Vit. D (10000 IU) were administered in all groups except the control. The number of muscle and inflammatory cells, fiber diameter, endomysium thickness, and degenerative collagen fiber area were assessed through hematoxylin-eosin staining.

Results: Muscle cell number was increased in the Ex + Vit. D + Ca, Vit. D + Ex, and Vit. D groups compared to the control group; also, inflammatory cell number showed significant increase in the Ex + Vit. D + Ca (12 ± 5.46), Vit. D + Ex (14 ± 3.25), Ex (13 ± 4.08), Vit. D (11 ± 3.26), Ca + Vit. D (10 ± 1.01), and Ca + Ex (9 ± 2.87) groups. Muscle fiber diameter in the Ex + Vit. D + Ca and Vit. D + Ex groups was higher than the other groups. Endomysium thickness was significantly decreased in the Ex + Vit. D + Ca and Vit. D + Ex groups compared to the control and placebo groups (p < 0.001). Degenerative collagen fiber area showed a significant increase in the Ex + Vit. D + Ca and Vit. D + Ex groups (p ≤ 0.001) comparison with the control group.

Conclusion: Regular resistance exercise, Vit. D, and Ca supplements can improve muscle morphological features in the postmenopausal period.

Key words: Menopause, Muscle, Vitamin D, Calcium, Exercise.

[1] Pace DT. The menopausal woman: The need for an individualized plan of care. Nurse Pract 2017; 42: 43–49.

[2] Nguyen HH, Milat F, Vincent A. Premature ovarian insufficiency in general practice: Meeting the needs of women. Aust Fam Physician 2017; 46: 360–366.

[3] Searles S, Makarewicz JA, Dumas JA. The role of estradiol in schizophrenia diagnosis and symptoms in postmenopausal women. Schizophr Res 2018; 196: 35–38.

[4] Larsson L, Degens H, Li M, Salviati L, Lee YI, Thompson W, et al. Sarcopenia: Aging-related loss of muscle mass and function. Physiol Rev 2019; 99: 427–511.

[5] Padilla Colon CJ, Molina-Vicenty IL, Frontera-Rodriguez M, Garcia-Ferre A, Rivera BP, Cintron-Velez G, et al. Muscle and bone mass loss in the elderly population: Advances in diagnosis and treatment. J Biomed 2018; 3: 40–49.

[6] Gimigliano F, Moretti A, de Sire A, Calafiore D, Iolascon G. The combination of vitamin D deficiency and overweight affects muscle mass and function in older post-menopausal women. Aging Clin Exp Res 2018; 30: 625–631.

[7] Kim JS, Jeon J, An JJ, Yi HK. Interval running training improves age-related skeletal muscle wasting and bone loss: Experiments with ovariectomized rats. Exp Physiol 2019; 104: 691–703.

[8] Estebanez B, de Paz JA, Cuevas MJ, Gonzalez-Gallego J. Endoplasmic reticulum unfolded protein response, aging and exercise: An update. Front Physiol 2018; 9: 1744.

[9] Kwon DH, Park HA, Cho YG, Kim KW, Kim NH. Different associations of socioeconomic status on protein intake in the korean elderly population: A cross-sectional analysis of the korea national health and nutrition examination survey. Nutrients 2020; 12: 10.

[10] Liberman K, Forti LN, Beyer I, Bautmans I. The effects of exercise on muscle strength, body composition, physical functioning and the inflammatory profile of older adults: A systematic review. Curr Opin Clin Nutr Metab Care 2017; 20: 30–53.

[11] Cheung MM, DeLuccia R, Ramadoss RK, Aljahdali A, Volpe SL, Shewokis PA, et al. Low dietary magnesium intake alters vitamin D-parathyroid hormone relationship in adults who are overweight or obese. Nutr Res 2019; 69: 82–93.

[12] Kruger MC, Chan YM, Lau LT, Lau CC, Chin YS, Kuhn- Sherlock B, et al. Calcium and vitamin D fortified milk reduces bone turnover and improves bone density in postmenopausal women over 1 year. Eur J Nutr 2018; 57: 2785–2794.

[13] Heshmat R, Mohammad K, Majdzadeh SR, Forouzanfar MH, Bahrami A, Ranjbar Omrani GH, et al. Vitamin D deficiency in Iran: A multi-center study among different urban areas. Iranian J Public Health 2008; 37: 72–78.

[14] Tabrizi R, Moosazadeh M, Akbari M, Dabbaghmanesh MH, Mohamadkhani M, Asemi Z, et al. High prevalence of vitamin D deficiency among Iranian population: A systematic review and meta-analysis. Iran J Med Sci 2018; 43: 125–139.

[15] Saki F, Dabbaghmanesh MH, Omrani GR, Bakhshayeshkaram M. Vitamin D deficiency and its associated risk factors in children and adolescents in southern Iran. Public Health Nutr 2017; 20: 1851–1856.

[16] Chen C, Noland KA, Kalu DN. Modulation of intestinal vitamin D receptor by ovariectomy, estrogen and growth hormone. Mech Age Dev 1997; 99: 109–122.

[17] Prestes J, Leite R, Pereira G, Shiguemoto G, Bernardes C, Asano R, et al. Resistance training and glycogen content in ovariectomized rats. Int J Sports Med 2012; 33: 550–554.

[18] Barzegar MH, Khazali H, Kalantar SM, Khoradmehr A. Effect of Citrullus colocynthis hydro-alcoholic extract on hormonal and folliculogenesis process in estradiol valerate-induced PCOs rats model: An experimental study. Int J Reprod Biomed 2017; 15: 661–668.

[19] Ruegg MA, Meinen S. Histopathology in hematoxylin & eosin stained muscle sections. Sop no MDC1A_M. 2014;1(004).

[20] Taebi M, Abdolahian S, Ozgoli G, Ebadi A, Kariman N. Strategies to improve menopausal quality of life: A systematic review. J Educ Health Promot 2018; 7: 93.

[21] Traish AM, Vignozzi L, Simon JA, Goldstein I, Kim NN. Role of androgens in female genitourinary tissue structure and function: Implications in the genitourinary syndrome of menopause. Sex Med Rev 2018; 6: 558–571.

[22] Maltais ML, Desroches J, Dionne IJ. Changes in muscle mass and strength after menopause. J Musculoskelet Neuronal Interact 2009; 9: 186–197.

[23] Fu S, Choy NL, Nitz J. Controlling balance decline across the menopause using a balance-strategy training program: A randomized, controlled trial. Climacteric 2009; 12: 165– 176.

[24] Hevener AL, Zhou Z, Drew BG, Ribas V. The role of skeletal muscle estrogen receptors in metabolic homeostasis and insulin sensitivity. Adv Exp Med Biol 2017; 1043: 257–284.

[25] Dehaini H, Fardoun M, Abou-Saleh H, El-Yazbi A, Eid AA, Eid AH. Estrogen in vascular smooth muscle cells: A friend or a foe? Vascul Pharmacol 2018; 111: 15–21.

[26] Abiri B, Vafa M. Vitamin D and muscle sarcopenia in aging. Methods Mol Biol 2020; 2138: 29–47.

[27] Yamada S, Tsuruya K, Yoshida H, Tokumoto M, Ueki K, Ooboshi H, et al. Factors associated with the serum myostatin level in patients undergoing peritoneal dialysis: Potential effects of skeletal muscle mass and vitamin D receptor activator use. Calcif Tissue Int 2016; 99: 13–22.

[28] Abrigo J, Simon F, Cabrera D, Cordova G, Trollet C, Cabello-Verrugio C. Central role of transforming growth factor type beta 1 in skeletal muscle dysfunctions: An update on therapeutic strategies. Curr Protein Pept Sci 2018; 19: 1189–1200.

[29] Latres E, Pangilinan J, Miloscio L, Bauerlein R, Na E, Potocky TB, et al. Myostatin blockade with a fully human monoclonal antibody induces muscle hypertrophy and reverses muscle atrophy in young and aged mice. Skelet Muscle 2015; 5: 34.

[30] Kim JS, Cross JM, Bamman MM. Impact of resistance loading on myostatin expression and cell cycle regulation in young and older men and women. Am J Physiol Endocrinol Metab 2005; 288: E1110–E1119.

[31] Montenegro KR, Cruzat V, Carlessi R, Newsholme P. Mechanisms of vitamin D action in skeletal muscle. Nutr Res Rev 2019; 32: 192–204.

[32] Phillips MD, Patrizi RM, Cheek DJ, Wooten JS, Barbee JJ, Mitchell JB. Resistance training reduces subclinical inflammation in obese, postmenopausal women. Med Sci Sports Exerc 2012; 44: 2099–2110.

[33] Sirola J, Rikkonen T. Muscle performance after the menopause. J Br Menopause Soc 2005; 11: 45–50.

[34] Grumati P, Coletto L, Schiavinato A, Castagnaro S, Bertaggia E, Sandri M, et al. Physical exercise stimulates autophagy in normal skeletal muscles but is detrimental for collagen VI-deficient muscles. Autophagy 2011; 7: 1415–1423.

[35] Mason C, Foster-Schubert KE, Imayama I, Kong A, Xiao L, Bain C, et al. Dietary weight loss and exercise effects on insulin resistance in postmenopausal women. Am J Prev Med 2011; 41: 366–375.

[36] Ortmeyer HK, Goldberg AP, Ryan AS. Exercise with weight loss improves adipose tissue and skeletal muscle markers of fatty acid metabolism in postmenopausal women. Obesity 2017; 25: 1246–1253.

[37] Joseph LJ, Davey SL, Evans WJ, Campbell WW. Differential effect of resistance training on the body composition and lipoprotein-lipid profile in older men and women. Metabolism 1999; 48: 1474–1480.

[38] Gries KJ, Minchev K, Raue U, Grosicki GJ, Begue G, Finch WH, et al. Single-muscle fiber contractile properties in lifelong aerobic exercising women. J Appl Physiol 2019; 127: 1710–1719.

[39] Fluck M, Kramer M, Fitze DP, Kasper S, Franchi MV, Valdivieso P. Cellular aspects of muscle specialization demonstrate genotype - phenotype interaction effects in athletes. Front Physiol 2019; 10: 526.

[40] Chambers TL, Burnett TR, Raue U, Lee GA, Finch WH, Graham BM, et al. Skeletal muscle size, function, and adiposity with lifelong aerobic exercise. J Appl Physiol 2020; 128: 368–378.

[41] Winnard A, Scott J, Waters N, Vance M, Caplan N. Effect of time on human muscle outcomes during simulated microgravity exposure without countermeasuressystematic review. Front Physiol 2019; 10: 1046.

[42] Consitt LA, Copeland JL, Tremblay MS. Endogenous anabolic hormone responses to endurance versus resistance exercise and training in women. Sports Med 2002; 32: 1–22.