?Metformin acts directly on rat granulosa cells activating ampk and regulating vegf expression

PARBORELL FERNANDA; OUBIÑA G,; DI PIETRO M; MARTA TESONE; PASCUALI N; ABRAMOVICH DAHIA; FRUNGIERI MB; L SCOTTI

Buenos Aires

Congreso; Reunión de las sociedades de Biociencias; 2017

Sociedades Argentinas de Biociencias

Polycystic ovary syndrome (PCOS) is a common disorder that affects women in reproductive age. Its symptoms are heterogeneous and range from anovulation, oligo/amenorrhea and hyperandrogenism to obesity and insulin resistance. Metformin (MET) is an oral antihyperglycemic drug introduced in the treatment of PCOS to manage hyperglycemia. MET has been shown to improve ovulation, pregnancy and live birth rates in patients with PCOS. The mechanism by which MET improves reproductive parameters are not fully understood. MET is a hydrophilic molecule so the organic cation transporters (OCTs) are actively involved in the celular uptake of MET in different tissues. MET primary mechanism of action is through the activation of the AMP-activated protein kinase (AMPK), which acts as an energy sensor by monitoring the AMP/ATP status of the cell. The aims of the present work were to analyze a possible direct effect of MET on rat granulosa cells (rGCs) and to evaluate the presence of OCTs in this cell type. Materials and Methods: Sprague Dawley rats (21d) were injected subcutaneously with diethylstilbestrol (1mg/rat) daily for three days to stimulate the development of early antral follicles. rGCs were isolated by percoll gradient. RNA was isolated from the rGCs and RT-PCR was performed for OCT 1-3. Another set of isolated rGCs was stimulated with MET 0.01 or 0.1ng/ml with or without the OCT inhibitor cimetidine (CIM). Cells were harvested 48h later and proteins extracted for OCTs, phospho-AMPK (P-AMPK), AMPK and VEGF measurement by western blot. Results: Expression of OCT 1-3 was detected in rGCs. P-AMPK was increased (P