In vivo, in vitro, and in silico studies of Cu/Zn-superoxide dismutase regulation by molecules in grape seed procyanidin extract.

The potential beneficial effects of flavonoids on human health have aroused considerable interest and were initially attributed to their antioxidant activities. Recent studies have speculated that as well as their antioxidant role, flavonoids can act by modulating cell signaling pathways and/or gene expression. In this respect, we have used streptozotocin-induced diabetic rats as an oxidative stress model to study whether grape seed procyanidin extract (GSPE) regulates copper/zinc-superoxide dismutase (Cu/Zn-SOD), an enzyme that defends against oxidative stress. The results indicate that the expression profile of Cu/Zn-SOD in diabetic rats was similar to the profile in nondiabetic rats. Nevertheless, the administration of GSPE increased Cu/Zn-SOD activity in both diabetic and nondiabetic rats. Therefore, to evaluate whether this increase in activity was dose-dependent, we also studied the effect of GSPE on Cu/Zn-SOD expression by using an in vitro model (Fao cell line hepatocytes). The cells were exposed to GSPE doses between 0 and 150 mg/L for 24 h, and the results showed that enzyme activity was enhanced only with 15 mg/L of GSPE. Therefore, we decided to explore whether this increase in Cu/Zn-SOD activity was due to direct interaction between some of the molecules in GSPE and the enzyme (in vitro experiments) and, if so, to analyze how this interaction occurs (in silico experiments). The results of these studies showed that direct interaction between some small- or medium-sized GSPE components and the enzyme is responsible for the increase in Cu/Zn-SOD activity.