Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus

Authors

DOI:

https://doi.org/10.33448/rsd-v10i6.15536

Keywords:

Interstitial cells; Ovarian follicles; Metformin; Rats.

Abstract

Objective: To evaluate the histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus. Methods: Thirty-two adult-female rats with regular estrous cycle were equally divided into four groups: 1) GCtrl - at estrous phase. 2) GPCOS - at permanent-estrous phase. 3) GMet1 - rats and daily treated with metformin (12.5 mg/Kg) during 60 consecutive days, as preventive form and 4) GMet2 - PCOS rats, which remained exposed to 60 days of continuous illumination and treated with metformin. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 (Casp-3) detections. Results: The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in Mel-treated groups, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells. The presence of corpus luteum along with an increase in the number of primary follicles in the Met2 group were noticed (p<0,01). A significant reduction in number of cysts and in the area occupied by interstitial cells, as well as a decrease in nuclear volume of interstitial cells, were noticed in the Met-treated groups, mainly in the Met2 group. The percentage of cell proliferation was significantly higher in granulosa cells of the Met-treated groups than PCOS group, mainly in the GMet2 (p<0,01), which was similar to the GCtrl group. On the other hand, the percentage of apoptosis (cleaved-caspase-3- positive cells) was significantly higher in the granulosa cells of GPCOS and Met-treated groups than the GCtrl group, but without significant difference, which showed weak cleaved caspase-3 immunoreactivity in those cells. Conclusion: The ovaries of rats treated with metformin showed a decrease in nuclear volume and in the area occupied by interstitial cells, presence of corpus luteum, in addition to a decrease in the number of cysts.

References

Batista, J. G., Soares-Jr, J. M., Maganhin, C. C., Simões, R. S., Tomaz, G., & Baracat, E. C. (2012). Assessing the benefits of rosiglitazone in women with polycystic ovary syndrome through its effects on insulin-like growth factor 1, insulin-like growth factor-binding protein-3 and insulin resistance: a pilot study. Clinics. 67(3):283-7. 10.6061/clinics/2012(03)14.

Ding, L., Gao, F., Zhang, M., Yan, W., Tang, R., & Zhang, C. J. (2016). Higher PDCD4 expression is associated with obesity, insulin resistance, lipid metabolism disorders, and granulosa cell apoptosis in polycystic ovary syndrome. Fertil Steril. 105(5):1330-1337.

Dunaif, A. (1997). Insulin resistance and the polylistic ovary syndrome: mechanism and implications for pathogenesis. Endocr. Rev. 18:774-800.

Goodarzi, M. O., Dumesic, D. A., Chazenbalk, G., & Azziz, R. (2011). Polycystic ovary syndrome: etiology, pathogenesis and diagnosis. Nat Rev Endocrinol. 7(4):219-31. 10.1038/nrendo.2010.217.

Lauretta, R., Lanzolla, G., Vici, P., Mariani, L., Moretti, C., & Appetecchia, M. (2016). Insulin-Sensitizers, Polycystic Ovary Syndrome and Gynaecological Cancer Risk. Int J Endocrinol. 2016:8671762.

Lombardi, L. A., Simões, R. S., Maganhin, C. C., da Silva, C. F., Maciel, G. A., Baracat, E. C., et al. (2012). Morphology of the interstitial cells of rat polycystic ovaries: an experimental study. Rev Bras Ginecol Obstet. 34(7):323-8. 0.1590/S0100-72032012000700006.

Lombardi, L. A., Simões, R. S., Maganhin, C. C., Baracat, M. C., Silva-Sasso, G. R., Florencio-Silva, R., Soares Jr, J. M., & Baracat, E. C. (2014). Immunohistochemical evaluation of proliferation, apoptosis and steroidogenic enzymes in the ovary of rats with polycystic ovary. Rev Assoc Med Bras. 60(4):349-56.

Maciel, G. A., Soares Jr, J. M., Motta, E. L. A., & Baracat, E. C. (2004). Nonobese women with polycystic ovary syndrome respond better than obese women to treatment with metformin. Fertil Steril. 81:355-60.

Mahamed, R. R., Maganhin, C. C., Simões, R. S., De Jesus Simões, M., Baracat, E. C., & Soares, J. M. Jr. (2011). Effects of metformin on the reproductive system of androgenized female rats. Fertil Steril. 95(4):1507-9.

Mathur, R., Alexander, C. J., Yano, J., Trivax, B., Azziz, R. (2008). Use of metformin in polycystic ovary syndrome. Am J Obstet Gynecol Reviews. 596-609.

Moll, E., Korevaar, J., Bossuyt, P. & Van Der Veen, F. (2008). Does adding metformin to clomifene citrate lead to higher pregnancy rates in a subset of women with polycystic ovary syndrome? Human Reproduction. 23(8), 1830-1834.

Nestler, J. E., Jakubowicz, D. J., Evans, W. S., & Pasquali, R. (1998). Effects of metformin on spontaneous and clomiphene-induced ovulation in the polycystic ovary syndrome. N Engl J Med. 338:1876-80.

Olfert, E. D., Cross, B. N., & Mcwilliam, A. A. (1993). Canadian Council on Animal Care/Guide to the Care and Use of Experimental Animals. 2nd ed. Ottawa, Ontario, Canada: Bradda Printing Services. https://www.ccac.ca/Documents/Standards/Guidelines/Experimental_Animals_Vol1.pdf.

Palomba, S., Orio, F. Jr, Falbo, A., Manguso, F., Russo, T., Cascella, T., Tolino, A., Carmina, E., Colao, A., Zullo, F. (2005). Prospective parallel randomized, doubleblind, double-dummy controlled clinicaltrial comparing clomiphene citrate and metformin as the first-line treatment for ovulation induction in non obese anovulatory women with polycystic ovary syndrome. J Clin Endocrinol Metab. 90:4068-74.

Penna, I. (2005). Acarbose in obese patients with polycystic ovarian syndrome: a double-blind, randomized,placebo-controlled study. Human Reproductio. 20(9), 2396-2401.

Qiao, J., Wang, L., Li, R., & Zhang, X. (2008). Microarray evaluation of endometrial receptivity in Chinese women with polycystic ovary syndrome. Reprod Biomed Online. 17(3):425-35.

Sara, L., Antal, P., Masszi, G., Buday, A., Horvath, E. M., Hamar, P., et al. (2012). Arteriolar insulin resistance in a rat model of polycystic ovary syndrome. Fertil Steril. 97(2):462-8. 10.1016/j.fertnstert.2011.11.015.

Smaniotto, S., & Simões, M. J. (1996). Morphological aspects of the endometrial epithelium of persistent estrous rats under estrogenic action. Braz J Morphol Sci. 13:103.

Speroff, L., & Fritz, M. A. (2005). Obesity. The physiology of adipose tissue, and the problem of obesity. In: Speroff L, Fritz MA, editors. Clinical gynecologic endocrinology and infertility. Philadelphia: Lippincott Williams & Wilkins. 779-804.

Tamimi, W., Siddiqui, Ia, Tamim, H., Aleisa, N., & Adham, M. (2009). Effect of body mass index on clinical manifestations in patients with polycystic ovary syndrome. Int J Gynaecol Obstet. 107(1):54-7.

Tang, T., Lord, J. M., Norman, R. J., Yasmin, E., & Balen, A. H. (2012). Insulin-sensitising drugs (metformin,rosiglitazone, pioglitazone, D-chiro-inositol) for women with polycystic ovary syndrome, oligo amenorrhoea and subfertility. Cochrane Database Syst Rev. (5):CD003053.

Vandermolen, D. T., Ratts, V. S., Evans, W. S., Stovall, D. W., Kauma, S. W., & Nestler, J. E. (2001). Metformin increases the ovulatory rate and pregnancy rate from clomiphene citrate in patients with polycystic ovary syndrome who are resistant to clomiphene citrate alone. Fertil Steril. 75:310-5.

Zabuliene, L., & Tutkuviene, J. (2010). Body composition and polycystic ovary syndrome. Medicina (Kaunas), 46(2):142-57.

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Published

20/05/2021

How to Cite

LOMBARDI, L. A. .; MATTOS, L. S. .; MOURA, M. L. A. .; ESPINDULA, A. P. .; SIMÕES, R. S. .; FLORENCIO-SILVA, R. .; SIMÕES, M. de J.; FRANCO, P. C. .; SASSO, G. R. da S.; SOARES-JR, J. M. . Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus . Research, Society and Development, [S. l.], v. 10, n. 6, p. e5110615536, 2021. DOI: 10.33448/rsd-v10i6.15536. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/15536. Acesso em: 20 jun. 2021.

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Health Sciences