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Journal of Ovarian Research
Published in: Journal of Ovarian Research 1/2018

Open Access 01-12-2018 | Research

Effect of resveratrol and metformin on ovarian reserve and ultrastructure in PCOS: an experimental study

Authors: Selenay Furat Rencber, Sema Kurnaz Ozbek, Ceyla Eraldemır, Zehra Sezer, Tugba Kum, Sureyya Ceylan, Elif Guzel

Published in: Journal of Ovarian Research | Issue 1/2018

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Abstract

Background

PCOS is a reproductive hormonal abnormality and a metabolic disorder. It is frequently associated with insulin resistance, hyperandrogenism, chronic inflammation, and oxidative stress. We aim to investigate the potential therapeutic effects of combined therapy of resveratrol and metformin on polycystic ovaries via SIRT1 and AMPK activation.

Methods

Wistar albino rats were divided into control and experimental (PCOS) groups. DHEA-induced PCOS rats were given resveratrol (20 mg/kg/day), metformin (300 mg/kg/day) and combined therapy. At the end of the experiment, the body and ovarian weight of rats were measured and blood samples were analyzed for FSH, LH, testosterone, AMH, TNF-α and MDA levels. Histopathological evaluation of ovaries were carried out by light and electron microscopy. SIRT1 and AMPK immunreactivity and TUNEL assay were scored. Data were statistically analyzed by SPSS programme.

Results

Metformin and combined treatment groups reduced the body and ovary weights compared to the PCOS group. Serum testosterone levels were significantly higher in the PCOS group than in the control group and this was reduced when PCOS was treated with all but especially resveratrol. All the treatment groups decreased LH, LH/FSH, TNF-α and tissue AMH levels which were induced in the PCOS group, whereas metformin was unable to improve the increased MDA and plasma AMH levels. Treatment with resveratrol and/or metformin ameliorated the elevated number of secondary and atretic follicles and the decreased number of Graafian follicles in the PCOS group, which indicates the effect of the treatments on the maintenance of folliculogenesis. Light and electron microscopic findings supported the analysis of follicular count. Increased number of TUNEL (+) granulosa cells in the PCOS group were reduced significantly in the treatment groups. Resveratrol and metformin increased SIRT1 and AMPK immunreactivity, respectively, compared to the PCOS group.

Conclusions

The results suggest that combined therapy of metformin and resveratrol may improve the weight gain, hormone profile and ovarian follicular cell architecture by inducing antioxidant and antiinflammatory systems via SIRT1 and AMPK activation in PCOS.
Literature
1.
Joham AE, Teede HJ, Ranasinha S, Zoungas S, Boyle J. Prevalence of infertility and use of fertility treatment in women with polycystic ovary syndrome: data from a large community-based cohort study. J Women's Health (Larchmt). 2015;24(4):299–307.CrossRef
2.
Murri M, Luque-ram’ırez M, Insenser MM, Ojeda-Ojeda M, Escobar-Morreale HF. Circulating markers of oxidative stress and polycystic ovary syndrome (PCOS): a systematic review and meta-analysis. Hum Reprod Update. 2013;19(3):268–88.CrossRefPubMed
3.
Zhang J, Fan P, Liu H, Bai H, Wang Y, Zhang F. Apolipoprotein A-I and B levels, dyslipidemia and metabolic syndrome in south-west Chinese women with PCOS. Hum Reprod. 2012;27(8):2484–93.CrossRefPubMed
4.
Ehrmann DA. Polycystic ovary syndrome. J Med. 2005;352:1223–36.
5.
Nisenblat V, Norman RJ. Androgens and polycystic ovary syndrome. Curr Opin Endocrinol, Diabetes Obes. 2009;16(3):224–31.CrossRef
6.
Spaczynski RZ, Arici A, Duleba AJ. Tumor necrosis factor-α stimulates proliferation of rat ovarian theca-interstitial cells. Biol Reprod 1999;61(4):993–998.CrossRefPubMed
7.
Repaci A, Gambineri A, Pasquali R. The role of low-grade inflammation in the polycystic ovary syndrome. Mol Cell Endocrinol. 2011;335(1):30–41.CrossRefPubMed
8.
Diamanti-Kandarakis E, Paterakis T, Alexandraki K, Piperi C, Aessopos A, Katsikis I, et al. Indices of low-grade chronic inflammation in polycystic ovary syndrome and the beneficial effect of metformin. Hum Reprod. 2006;21(6):1426–31.CrossRefPubMed
9.
Gonz’alez F, Rote NS, Minium J, Kirwan JP. Evidence of proatherogenic inflammation in polycystic ovary syndrome. Metab Clin Exp. 2009;58(7):954–62.CrossRef
10.
Gonz’alez F. Inflammation in polycystic ovary syndrome: underpinning of insulin resistance and ovarian dysfunction. Steroids. 2012;77(4):300–5.CrossRef
11.
Ghafurniyan H, Azarnia M, Nabiuni M, Karimzadeh L. The effect of green tea extract on reproductive improvement in estradiol valerate-induced polycystic ovarian syndrome in rat. Iran J Pharm Res. 2015;14(4):1215–33.PubMedPubMedCentral
12.
Parahuleva N, Pehlivanov B, Dimitrakova E, Malinova M, Mladenova M. Anti-Mullerian hormone- its role in the pathogenesis of the polycystic ovary syndrome. Akush Ginekol. 2012;51(6):22–6.
13.
Iliodromiti S, Kelsey TW, Anderson RA, Nelson SM. Can anti-Mullerian hormone predict the diagnosis of polycystic ovary syndrome? A systematic review and meta-analysis of extracted data. J Clin Endocrinol Metab. 2013;98(8):3332–40.CrossRefPubMed
14.
15.
Rege SD, Kumar S, Wilson DN, Tamura L, Geetha T, Mathews ST, et al. Resveratrol protects the brain of obese mice from oxidative damage. Oxidative Med Cell Longev. 2013;419092
16.
17.
Guo S, Yao Q, Ke Z, Chen H, Wu J, Liu C. Resveratrol attenuates high glucose-induced oxidative stress and cardiomyocyte apoptosis through AMPK. Mol Cell Endocrinol. 2015;412:85–94.CrossRefPubMed
18.
Harati K, Slodnik P, Chromik AM, Goertz O, Hirsch T, Kapalschinski N, et al. Resveratrol induces apoptosis and alters gene expression in human fibrosarcoma cells. Anticancer Res. 2015;35(2):767–74.PubMed
19.
Wu XP, Xiong M, Xu CS, Duan LN, Dong YQ, Luo Y, et al. Resveratrol induces apoptosis of human chronic myelogenous leukemia cells in vitro through p38 and JNK-regulated H2AX phosphorylation. Acta Pharma. 2015;36(3):353–61.CrossRef
20.
Singh CK, Kumar A, Hitchcock DB, Fan D, Goodwin R, LaVoie HA, et al. Resveratrol prevents embryonic oxidative stress and apoptosis associated with diabetic embryopathy and improves glucose and lipid profile of diabetic dam. Mol Nutr Food Res. 2011;55(8):1186–96.CrossRefPubMedPubMedCentral
21.
MacCarrone M, Lorenzon T, Guerrieri P, Agrô AF. Resveratrol prevents apoptosis in K562 cells by inhibiting lipoxygenase and cyclooxygenase activity. Eur J Biochem. 1999;265(1):27–34.CrossRefPubMed
22.
Kong XX, Fu YC, Xu JJ, Zhuang XL, Chen ZG, Luo LL. Resveratrol, an effective regulator of ovarian development and oocyte apoptosis. J Endocrinol Investig. 2011;34(11):374–81.
23.
Liu Y, He XQ, Huang X, Ding L, Xu L, Shen YT, et al. Resveratrol protects mouse oocytes from methylglyoxal-induced oxidative damage. PLoS One. 2013;23;8(10):e77960.CrossRef
24.
Borra MT, Smith BC, Denu JM. Mechanism of human SIRT1 activation by resveratrol. J Biol Chem. 2005;280(17):17187–95.CrossRefPubMed
25.
Wilking MJ, Singh C, Nihal M, Zhong W, Ahmad N. SIRT1 deacetylase is overexpressed in human melanoma and its small molecule inhibition imparts anti-proliferative response via p53 activation. Arch Biochem Biophys. 2014;563:94–100.CrossRefPubMedPubMedCentral
26.
Lee D, Goldberg AL. SIRT1 protein, by blocking the activities of transcription factors FoxO1 and FoxO3, inhibits muscle atrophy and promotes muscle growth. J Biol Chem. 2013;288:30515–26.CrossRefPubMedPubMedCentral
27.
Tatone C, Di Emidio G, Vitti M, Di Carlo M, Santini S Jr, D'Alessandro AM, et al. Sirtuin functions in female fertility: possible role in oxidative stress and aging. Oxidative Med Cell Longev. 2015;2015:659687.CrossRef
28.
Kai Y, Kawano Y, Yamamoto H, Narahara H. A possible role for AMP-activated protein kinase activated by metformin and AICAR in human granulosa cells. Reprod Biol Endocrinol. 2015;13(1):1.CrossRef
29.
Patel R, Shah G. Effect of metformin on clinical, metabolic and endocrine outcomes in women with polycystic ovary syndrome: a metaanalysis of randomized controlled trials. Curr Med Res Opin. 2017:1–13.
30.
Ladson G, Dodson WC, Sweet SD, Archibong AE, Kunselman AR, Demers LM, et al. The effects of metformin with lifestyle therapy in polycystic ovary syndrome: a randomized double blind study. Fertil Steril. 2011;95(3):1059–66.CrossRefPubMed
31.
Palomba S, Orio F Jr, Falbo A, Manguso F, Russo T, Cascella T, et al. Prospective parallel randomized, double-blind, double-dummy controlled clinical trial 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. 2005;90(7):4068–74.CrossRefPubMed
32.
Attia GR, Rainey WE, Carr BR. Metformin directly inhibits androgen production in human thecal cells. Fertil Steril. 2001;76(3):517–24.CrossRefPubMed
33.
Mansfield R, Galea R, Brincat M, Hole D, Mason H. Metformin has direct effects on human ovarian steroidogenesis. Fertil Steril. 2003;79(4):956–62.CrossRefPubMed
34.
Corton JM, Gillespie JG, Hardie DG. Role of the AMP-activated protein kinase in the cellular stress response. Curr Biol. 1994;4:315–24.CrossRefPubMed
35.
Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, et al. Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest. 2001;108(8):1167–74.CrossRefPubMedPubMedCentral
36.
Salminen A, Hyttinen JM, Kaarniranta K. AMP-activated protein kinase inhibits NF-kappa B signaling and inflammation: impact on healthspan and lifespan. J Mol Med (Berl). 2011;89(7):667–76.CrossRef
37.
Shin SM, Cho IJ, Kim SG. Resveratrol protects mitochondria against oxidative stress through AMP-activated protein kinase-mediated glycogen synthase kinase-3 inhibition downstream of poly(ADP-ribose) polymerase-LKB1 pathway. Mol Pharmacol. 2009;76(4):884–95.CrossRefPubMed
38.
Okayasu S, Kitaichi K, Hori A, Suwa T, Horikawa Y, Yamamoto M, et al. The evaluation of risk factors associated with adverse drug reactions by metformin in type 2 diabetes mellitus. Biol Pharm Bull. 2012;35(6):933–7.CrossRefPubMed
39.
Zhang X, Zhang C, Shen S, Yj X, Yi L, Gao Q, et al. Dehydroepiandrosterone induces ovarian and uterine hyperfibrosis in female rats. Hum Reprod. 2013;28(11):3074–85.CrossRefPubMed
40.
Erdem T, Bayindir T, Filiz A, Iraz M, Selimoglu E. The effect of resveratrol on the prevention of cisplatin ototoxicity. Eur Arch Otorhinolaryngol. 2012;269:2185–8.CrossRefPubMed
41.
Reagan-Shaw S, Nihal M, Ahmad N. Dose translation from animal to human studies revisited. FASEB J. 2008;22:659–61.CrossRefPubMed
42.
Calkins CM, Bensard DD, Heimbach JK, Meng X, Shames BD, Pulido EJ, et al. L-arginine attenuates lipopolysaccharide induced lung chemokine production. Am. J. Physiol. Lung Cell Mol. Physiol. 2001;280(3):L400–8.CrossRefPubMed
43.
Hartree EF. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972;48:422–7.CrossRefPubMed
44.
45.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis. 2005;15(4):316–28.CrossRefPubMed
46.
Dorostghoal M, Mahabadi MK, Adham S. Effects of maternal caffeine consumption on ovarian follicle development in wistar rats offsping. J Reprod Infertil. 2011;12(1):15–22.PubMedPubMedCentral
47.
Atilgan R, Kuloglu T, Boztosun A, Orak U, Baspinar M, Can B, et al. Investigation of the effects of unilateral total salpingectomy on ovarian proliferating cell nuclear antigen and follicular reserve: experimental study. Eur J Obstet Gynecol Reprod Biol. 2015;188:56–60.CrossRefPubMed
48.
Medigović I, Ristić N, Trifunović S, Manojlović-Stojanoski M, Milošević V, Zikić D, et al. Genistein affects ovarian folliculogenesis: a stereological study. Microsc Res Tec. 2012;75(12):1691–9.CrossRef
49.
Tao X, Zhang X, Ge S, Zhang E, Zhang B. Expression of SIRT1 in the ovaries of rats with polycystic ovary syndrome before and after therapeutic intervention with exenatide. Int J Clin Exp Pathol. 2015;8(7):8276–83.PubMedPubMedCentral
50.
Ma MQ, Thapalia BA, Lin XH. A 6 hour therapeutic window, optimal for interventions targeting AMPK synergism and apoptosis antagonism, for cardioprotection against myocardial ischemic injury: an experimental study on rats. Am J Cardiovasc Dis. 2015;5(1):63–71.PubMedPubMedCentral
51.
Guzel E, Basar M, Ocak N, Arici A, Kayisli UA. Bidirectional interaction between unfolded-protein-response key protein HSPA5 and estrogen signaling in human endometrium. Biol Reprod. 2011;85(1):121–7.CrossRefPubMed
52.
Kilic S, Yuksel B, Lortlar N, Sertyel S, Aksu T, Batioglu S. Environmental tobacco smoke exposure during intrauterine period promotes granulosa cell apoptosis: a prospective, randomized study. J Matern Fetal Neonatal Med. 2012;25(10):1904–8.CrossRefPubMed
53.
Sun J, Zhao JM, Ji R, Liu HR, Shi Y, Jin CL. Effects of electroacupuncture of "Guanyuan" (CV 4)-"Zhongji" (CV 3) on ovarian P450 arom and P450c 17alpha expression and relevant sex hormone levels in rats with polycystic ovary syndrome. Zhen Ci Yan Jiu. 2013;38(6):465–72.PubMed
54.
Wang F, Yu B, Yang W, Liu J, Lu J, Xia X. Polycystic ovary syndrome resembling histopathological alterations in ovaries from prenatal androgenized female rats. J Ovarian Res. 2012;18:5(1):15.CrossRef
55.
Walters KA, Allan CM, Handelsman DJ. Rodent models for human polycystic ovary syndrome. Biol Reprod. 2012;86(5):149. 1–12CrossRefPubMed
56.
Misugi T, Ozaki K, El Beltagy K, Tokuyama O, Honda K, Ishiko O. Insulin-lowering agents inhibit synthesis of testosterone in ovaries of DHEA-induced PCOS rats. Gynecol Obstet Investig. 2006;61:208–15.CrossRef
57.
Motta AB. Dehydroepiandrosterone to induce murine models for the study of polycystic ovary syndrome. J Steroid Biochem. 2010;119:105–11.CrossRef
58.
Sun L, Ji C, Jin L, Bi Y, Feng W, Li P, et al. Effects of Exenatide on metabolic changes, sexual hormones, inflammatory cytokines, adipokines, and weight change in a DHEA-treated rat model. Reprod Sci. 2016;23(9):1242–9.CrossRefPubMed
59.
Zang M, Zuccollo A, Hou X, Nagata D, Walsh K, Herscovitz H, et al. AMP-activated protein kinase is required for the lipid-lowering effect of metformin in insulin-resistant human HepG2 cells. J Biol Chem. 2004;279(46):47898–905.CrossRefPubMed
60.
61.
Wong DH, Villanueva JA, Cress AB, Duleba AJ. Effects of resveratrol on proliferation and apoptosis in rat ovarian theca-interstitial cells. Mol Hum Reprod. 2010;16(4):251–9.CrossRefPubMedPubMedCentral
62.
Ortega I, Villanueva JA, Wong DH, Cress AB, Sokalska A, Stanley SD, et al. Resveratrol potentiates effects of simvastatin on inhibition of rat ovarian theca-interstitial cells steroidogenesis. J Ovarian Res. 2014;7(1):1.CrossRef
63.
Ortega I, Villanueva JA, Wong DH, Cress AB, Sokalska A, Stanley SD, et al. Resveratrol reduces steroidogenesis in rat ovarian theca-interstitial cells: the role of inhibition of Akt/ PKB signaling pathway. Endocrinol. 2012;153(8):4019–29.CrossRef
64.
Henry LA, Witt DM. Resveratrol: phytoestrogen effects on reproductive physiology and behavior in female rats. Horm Behav. 2002;41:220–8.CrossRefPubMed
65.
Stracquadanio M, Ciotta L, Palumbo MA. Relationship between serum anti-Mullerian hormone and intrafollicular AMH levels in PCOS women. Gynecol Endocrinol. 2017;23:1–6.
66.
Matsuda F, Inoue N, Manabe M, Ohkura S. Follicular growth and atresia in mammalian ovaries: regulation by survival and death of granulosa cells. J Reprod Dev. 2012;58(1):44–50.CrossRefPubMed
67.
Ergenoglu M, Yildirim N, Yildirim AG, Yeniel O, Erbas O, Yavasoglu A, et al. Effects of resveratrol on ovarian morphology, plasma anti-Mullerian hormone, IGF-1 levels, and oxidative stress parameters in a rat model of polycystic ovary syndrome. Reprod Sci. 2015;22(8):942–7.CrossRefPubMed
68.
69.
Banu SK, Stanley JA, Sivakumar KK, Arosh JA, Burghardt RC. Resveratrol protects the ovary against chromium-toxicity by enhancing endogenous antioxidant enzymes and inhibiting metabolic clearance of estradiol. Toxicol Appl Pharmacol. 2016;15(303):65–78.CrossRef
70.
Cheserek MJ, Wu G, Li L, Li L, Karangwa E, Shi Y, et al. Cardioprotective effects of lipoic acid, quercetin and resveratrol on oxidative stress related to thyroid hormone alterations in long-term obesity. J Nutr Biochem. 2016;33:36–44.CrossRefPubMed
71.
Gonza’lez F, Rote NS, Minium J, Kirwan JP. Increased activation of nuclear factor-κB triggers inflammation and insulin resistance in polycystic ovary syndrome. J Clin Endocrinol Metab 2006;91(4):1508–1512.
72.
Mohammadi S, Kayedpoor P, Karimzadeh-Bardei L, Nabiuni M. The effect of curcumin on TNF-α, IL-6 and CRP expression in a model of polycystic ovary syndrome as an inflammation state. J Reprod Infertil. 2017;18(4):352–60.PubMedPubMedCentral
73.
Taguchi A, Wada-Hiraike O, Kawana K, Koga K, Yamashita A, Shirane A, et al. Resveratrol suppresses inflammatory responses in endometrial stromal cells derived from endometriosis: a possible role of the sirtuin 1 pathway. Obstet Gynaecol Res. 2014;40(3):770–8.CrossRef
74.
Orio F, Manguso F, Di Biase S, Falbo A, Giallauria F, Labella D, et al. Metformin administration improves leukocyte count in women with polycystic ovary syndrome: a 6-month prospective study. Eur J Endocrinol. 2007;157(1):69–73.CrossRefPubMed
75.
Ortega I, Wong DH, Villanueva JA, Cress AB, Sokalska A, Stanley SD, et al. Effects of resveratrol on growth and function of rat ovarian granulosa cells. Fertil Steril. 2012;98(6):1563–73.CrossRefPubMedPubMedCentral
76.
Sun J, Jin C, Wu H, Zhao J, Cui Y, Liu H, et al. Effects of electro-acupuncture on ovarian P450arom, P450c17a and mRNA expression induced by letrozole in PCOS rats. PLoS One. 2013;8(11):e79382.CrossRefPubMedPubMedCentral
77.
Grossman MP, Nakajima ST, Fallat ME, Siow Y. Mullerian-inhibiting substance inhibits cytochrome P450 aromatase activity in human granulosa lutein cell culture. Fertil Steril. 2008;89:1364–70.CrossRefPubMed
78.
79.
Leonard SS, Xia C, Jiang BH, Stinefelt B, Klandorf H, Harris GK, et al. Resveratrol scavenges reactive oxygen species and effects radical-induced cellular responses. Biochem Biophys Res Commun. 2003;309:1017–26.CrossRefPubMed
80.
Wang S, Wang J, Zhao A, Li J. SIRT1 activation inhibits hyperglycemia-induced apoptosis by reducing oxidative stress and mitochondrial dysfunction in human endothelial cells. Mol Med Rep. 2017;16(3):3331–8.CrossRefPubMed
Metadata
Title
Effect of resveratrol and metformin on ovarian reserve and ultrastructure in PCOS: an experimental study
Authors
Selenay Furat Rencber
Sema Kurnaz Ozbek
Ceyla Eraldemır
Zehra Sezer
Tugba Kum
Sureyya Ceylan
Elif Guzel
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Journal of Ovarian Research / Issue 1/2018
Electronic ISSN: 1757-2215
DOI
https://doi.org/10.1186/s13048-018-0427-7

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