1 ECED/08-2013-0249-Endo/19.12.2013/MPS Article ECED/08-2013-0249-Endo/19.12.2013/MPS Dăneasă A et al. Spironolactone and Dimethylsulfoxide Effect … Exp Clin Endocrinol Diabetes 2014; 122: 1–9 ■ Proof copy for correction only. All forms of publication, duplication or distribution prohibited under copyright law. ■ received 16.08.2013 first decision 30.10.2013 accepted 06.12.2013 Bibliography DOI http://dx.doi.org/ 10.1055/s-0033-1363685 Exp Clin Endocrinol Diabetes 2014; 122: 1–9 © J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York ISSN 0947-7349 Correspondence Prof. G. A. Filip Physiology Iuliu Hatieganu University of Medicine and Pharmacy Clinicilor Street No 1 Cluj Napoca Romania 400006 Tel.: + 40/745/268 704 Fax: + 40/264/598 575 gabriela.filip@umfcluj.ro Key words ● ▶ polycystic ovarian syndrome ● ▶ fasting hyperglycaemia ● ▶ glucose transporter 4 ● ▶ oxidative stress ● ▶ spironolactone ● ▶ dimethylsulfoxide Spironolactone and Dimethylsulfoxide Effect on Glucose Metabolism and Oxidative Stress Markers in Polycystic Ovarian Syndrome Rat Model One Sentence Summary: The purpose of our study is to explore the effect of antiandrogenic spironolactone (Sp) on glucose metabolism and oxidative stress in oestradiol valerate induced polycystic ovarian syndrome rat model. Sp in DMSO increased GLUT4 immunoreactivity and improved antioxidant capacity. environmental factors. For a positive diagnosis of PCOS 2 out of the following criteria are required: oligo and/or anovulation, clinical and/or bio- chemical signs of hyperandrogenism, and poly- cystic ovaries [3]. Besides these characteristics, around 60 % of women with PCOS present marked insulin resistance [4], which together with hyper- androgenism constitute the main pathological aspects of the syndrome. Moreover, PCOS is a risk factor for type 2 diabetes mellitus (T2DM) [5], the probability of developing frank diabetes being Introduction ▼ Polycystic ovarian syndrome (PCOS), the most common cause of infertility [1], affects 7–8 % of reproductive age women [2] being probably the most frequent endocrinopathy in women. PCOS definition remains an evolving and difficult task since its complex and incompletely under- stood pathophysiology results from the inter- action of endocrine, metabolic, genetic and Authors A. Dăneasă 1 *, C. Cucolaș 1 *, M. Furcea 1 , P. Bolfa 2 , S. Dudea 3 , D. Olteanu 1 , M. C. Alupei 4, 5 , A. Mureșan 1 , G. A. Filip 1 Abstract ▼ Because polycystic ovarian syndrome (PCOS) is a risk factor for type 2 diabetes, the affected women can present frequently prediabetic states such as impaired fasting glycaemia and/or impaired glucose tolerance. The purpose of our study is to explore the effect of antiandrogenic spironolactone on glucose metabolism and oxi- dative stress (OS) parameters in oestradiol valer- ate (OV) induced PCOS rat model. 72 female Wistar rats were distributed either to PCOS group (n = 65, OV dissolved in sesame oil, 5 mg/0.4 ml), or to non-PCOS control group (n = 7, sesame oil, 0.4 ml). After a month, ultrasound was performed to assess the ovarian morphol- ogy, and the results of an initial oral glucose tol- erance test (OGTT) were used to identify the animals with altered glucose metabolism (AGM). Glucose transporter 4 (GLUT4) was evaluated from muscle biopsies, OS parameters were assessed from blood and muscle samples, and ovaries of 3 rats were removed for histopatho- logical examination. Afterwards, the AGM group was divided in a treated PCOS group denoted as Sp + D (per os spironolactone dissolved in DMSO, 2 mg/0.2 ml), and a PCOS control treated with DMSO (0.2 ml). After one month of daily treat- ment, a final OGTT was performed. GLUT4 and OS parameters were again evaluated and ovaries were removed for histopathological examination. As compared to the values prior to the treatment, Sp + D reversed fasting hyperglycaemia (p < 0.001), increased GLUT4 immunoreactivity in the peri- nuclear compartment (p < 0.05) and transloca- tion to plasmalemma (p < 0.001) and improved superoxide dismutase (0.001 < p < 0.01) and glu- tathione peroxidase (0.001 < p < 0.01) activities, while reducing GSH level (0.001 < p < 0.01). Administration of DMSO alone decreased fasting hyperglycaemia (p < 0.001) and 2-h glucose level (p < 0.05) independently of GLUT4 translocation, improved superoxide dismutase (p < 0.001) and glutathione peroxidase (p < 0.05) activities in erythrocytes, reduced GSH level in serum (p < 0.05) and diminished lipid peroxidation in muscle as compared to the values recorded before treatment (0.001 < p < 0.01). Our results showed that the Sp + D treatment improved antioxidant capacity and had a benefi- cial effect on metabolic deregulation in PCOS. Administration of DMSO had an unexpected hypoglycaemiant effect and improved OS param- eters. This may represent an indirect proof of the role of oxidative stress and inflammation in PCOS and glucose metabolism abnormalities encoun- tered in PCOS. Affiliations Affiliation addresses are listed at the end of the article * These authors contributed equally to this work. two