In Vitro Effect of Resveratrol Supplementation on Oxidative Balance and Intercellular Communication of Leydig Cells Subjected to Induced Oxidative Stress

Abstract

Many studies have revealed that oxidative stress is a primary factor in the pathogenesis of male reproductive system dysfunctions. The strong antioxidant and cytoprotective effects of resveratrol have previously been demonstrated, but its effect in the context of the male reproduction remains unconvincing. To observe the biological activity of resveratrol in protecting the male reproductive function, hydrogen peroxide-induced oxidative stress in Leydig cells was used as a cell model. The aim of the present study was to examine if resveratrol could induce changes in the gap junction intercellular communication (GJIC), nitric oxide production, total oxidant status (TOS) and total antioxidant capacity (TAC) in TM3 Leydig cells subjected to H2O2. The Leydig cells were exposed to a resveratrol treatment (5, 10, 20, 50 and 100 μM) in the presence or absence of H2O2 (300/600 μM) during a 24 h in vitro culture. The cell lysates to assess TOS and TAC, NO production were quantified in a culture medium using the Griess method, and the Scrape Loading/Dye Transfer (SL/DT) technique was used for the determination of GJIC in the exposed TM3 Leydig cells. Treatment with higher doses of resveratrol alone led to a significantly increased TOS (p<0.05 with 100 μM) and NO production (p<0.05 with 50 μM and 100 μM), but significantly reduced TAC (p<0.01 with 100 μM) and GJIC (p<0.05 with 100 μM), while the SL/DT evaluation in the cells exposed to resveratrol at concentrations 5 μM (p<0.05) and 10 μM (p<0.01) revealed a significant stimulation of GJIC. The most potent cytoprotective or stimulatory effect of resveratrol in the cells co-exposed to oxidative stress (300 μM H2O2) was observed at a concentration of 10 μM in the case of GJIC, which was manifested by a significant increase in the values (p<0.05) compared to the control group treated with H2O2 alone.