The effects of metformin on stereological and ultrastructural features of the ovary in streptozotocin-induced diabetes adult rats: An experimental study

Abstract

Background: Diabetes is a chronic disease that can affect almost all of the body organs, including male and female reproductive systems.


Objective: This study was designed to investigate the preventive effects of metformin on stereological and ultrastructure characteristics of the ovary in the streptozotocin-induced diabetes adult female rats.


Materials and Methods: Seventy adult (8-10 wk) female Sprague-Dawley rats (180-200 gr) were equally divided, as follows: (n = 10/each) control; STZ-induced diabetes (single dose of 65 mg/kg STZ, IP); metformin-treated (50 mg/100 gr of body weight, orally); diabetic-metformin-treated; sham 1, (single dose of sodium citrate); sham 2, (0.5 ml of daily oral distilled water); and sham 3, (sodium citrate + distilled water treated). The body mass index, ovarian weight, blood sugar level, cholesterol, and triglyceride were measured. The stereological and ultrastructural features of ovary were assessed.


Results: The blood sugar of induced-diabetic rats was increased (p < 0.01). The BMI (p < 0.01), number of granulosa cells (p = 0.04), primordial, primary and secondary follicles (p = 0.03), total volume of ovary (p < 0.01) and cortex, nucleus diameter ratio to the ooplasm were decreased. The number of atretic follicles in the diabetic and diabetic + metformin-treated rats were increased (p < 0.01). The ultrastructural characteristics of ovary were more damaged in diabetic rats.


Conclusion: Diabetes has destructive effects on ovarian follicles and causes follicular atresia. Also, the size of oocytes, numbers of granulosa cells and ooplasmic organelles, which are involved in the folliculogenesis are affected by diabetes and metformin has no preventive effects.


Key words: Diabetes, Metformin, Ovary, Tissue.

References
[1] Whiting DR, Guariguata L, Weil C, Shaw J. IDF diabetes atlas: global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Res Clin Pract 2011; 94: 311–321.

[2] Wild S, Roglic G, Green A, Sicree R, King H. Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care 2004; 27: 1047–1053.

[3] Steger RW, Rabe MB. The effect of diabetes mellitus on endocrine and reproductive function. Proc Soc Exp Biol Med 1997; 214: 1–11.

[4] Chang AS, Dale AN, Moley KH. Maternal diabetes adversely affects preovulatory oocyte maturation, development, and granulosa cell apoptosis. Endocrinology 2005; 146: 2445–2453.

[5] Wu Y, Zhang Z, Liao X, Wang Z. High fat diet triggers cell cycle arrest and excessive apoptosis of granulosa cells during the follicular development. Biochem Biophys Res Commun 2015; 466: 599–605.

[6] Colton SA, Pieper GM, Downs SM. Altered meiotic regulation in oocytes from diabetic mice. Biol Reprod 2002; 67: 220–231.

[7] Chabrolle C, Jeanpierre E, Tosca L, Ramé C, Dupont J. Effects of high levels of glucose on the steroidogenesis and the expression of adiponectin receptors in rat ovarian cells. Reprod Biol Endocrinol 2008; 6: 11–24.

[8] Wu Y, Li Y, Liao X, Wang Z, Li R, Zou S, et al. Diabetes induces abnormal ovarian function via triggering apoptosis of granulosa cells and suppressing ovarian angiogenesis. Int J Biol Sci 2017; 13: 1297–1308.

[9] King AJ. The use of animal models in diabetes research. Br J Pharmacol 2012; 166: 877–894.

[10] Kruszyńska A, Słowińska-Srzednicka J, Jeske W, Zgliczyński W. Proinsulin, adiponectin and hsCRP in reproductive age women with polycystic ovary syndrome (PCOS)-the effect of metformin treatment. Endokrynol Pol 2014; 65: 2–10.

[11] Kennedy M, Dunning P. Diabetes education: Essential but underfunded in Australia. Diabetes and Primary Care Australia 2017; 2: 10–14.

[12] Diamanti−Kandarakis E, Economou F, Palimeri S, Christakou C. Metformin in polycystic ovary syndrome. Annals of the New York Academy of Sciences 2010; 1205: 192–198.

[13] Todd JN, Florez JC. An update on the pharmacogenomics of metformin: progress, problems and potential. Pharmacogenomics 2014; 15: 529–539.

[14] Elia E, Sander V, Luchetti CG, Solano ME, Di Girolamo G, Gonzalez C, et al. The mechanisms involved in the action of metformin in regulating ovarian function in hyperandrogenized mice. Mol Hum Reprod 2006; 12: 475– 481.

[15] Meng XM, Ma XX, Tian YL, Jiang Q, Wang LL, Shi R, et al. Metformin improves the glucose and lipid metabolism via influencing the level of serum total bile acids in rats with streptozotocin-induced type 2 diabetes mellitus. Eur Rev Med Pharmacol Sci 2017; 21: 2232–2237.

[16] Shirwaikar A, Rajendran K, Barik R. Effect of aqueous bark extract of Garuga pinnata Roxb. in streptozotocinnicotinamide induced type-II diabetes mellitus. J Ethnopharmacol 2006; 107: 285–290.

[17] Feldman M, Schiller LR. Disorders of gastrointestinal motility associated with diabetes mellitus. Ann Intern Med 1983; 98: 378–384.

[18] Akbarzadeh A, Norouzian D, Mehrabi MR, Jamshidi Sh, Farhangi A, Verdi AA, et al. Induction of diabetes by streptozotocin in rats. Indian J Clin Biochem 2007; 22: 60– 64.

[19] Courteix C, Eschalier A, Lavarenne J. Streptozocin-induced diabetic rats: behavioural evidence for a model of chronic pain. Pain 1993; 53: 81–88.

[20] Tesone M, Ladenheim RG, Oliveira-Filho RM, Chiauzzi VA, Foglia VG, Charreau EH. Ovarian dysfunction in streptozotocin-induced diabetic rats. Proc Soc Exp Biol Med 1983; 174: 123–130.

[21] Oppermann K, Fuchs SC, Spritzer PM. Ovarian volume in pre-and perimenopausal women: a population-based study. Menopause 2003; 10: 209–213.

[22] Eppig JJ. Mouse oocyte development in vitro with various culture systems. Dev Biol 1977; 60: 371–388.

[23] Erbas O, Pala HG, Pala EE, Oltulu F, Aktug H, Yavasoglu A, et al. Ovarian failure in diabetic rat model: Nuclear factorkappaB, oxidative stress, and pentraxin-3. Taiwan J Obstet Gynecol 2014; 53: 498–503.

[24] Sun QY, Nagai T. Molecular mechanisms underlying pig oocyte maturation and fertilization. J Reprod Dev 2003; 49: 347–359.

[25] Afrough M, Erfani Majd N, Najafzadeh H. Protective effects of aloe vera on superovulated oocytes and folliculogenesis in diabetic mice. Zahedan J Res Med Sci 2016; 18: e7939–e7942.

[26] Huang Z, Wells D. The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod 2010; 16: 715–725.

[27] Magoffin DA. Ovarian theca cell. Int J Biochem Cell Biol 2005; 37: 1344–1349.

[28] Wassarman P, Chen J, Cohen N, Litscher E, Liu C, Qi H, et al. Structure and function of the mammalian egg zona pellucida. J Exp Zool 1999; 285: 251–258.

[29] Bertrand E, Van den Bergh M, Englert Y. Fertilization and early embryology: does zona pellucida thickness influence the fertilization rate? Hum Reprod 1995; 10: 1189–1193.

[30] Szymanski KM, Binns D, Bartz R, Grishin NV, Li WP, Agarwal AK, et al. The lipodystrophy protein seipin is found at endoplasmic reticulum lipid droplet junctions and is important for droplet morphology. Proc Nati Acad Sci USA 2007; 104: 20890–20895.

[31] Pirwany IR, Yates RW, Cameron IT, Fleming R. Effects of the insulin sensitizing drug metformin on ovarian function, follicular growth and ovulation rate in obese women with oligomenorrhoea. Hum Reprod 1999; 14: 2963–2968.

[32] Barrett-Connor E, Khaw KT, Yen SSC. Endogenous sex hormone levels in older adult men with diabetes mellitus. Am J Epidemiol 1990; 132: 895–901.