Mijit Mahmut
L.Aquille University
Biography:
Abstract:
Abstract
Accumulation of Reactive Oxygen Species (ROS) and methlyglyoxal (MG) causes endothelial dysfunction in diabetes-related complications. Endothelial function in high glucose (HG) is improved by resveratrol (RSV), however the mechanisms underlying such beneficial effect are still unclear. In particular, the role of sirtuin-dependent regulation of cytoprotective mechanisms remains unknown.
Here we investigated molecular details glycative/oxidative perturbations in HG-challenged endothelial cells, and whether SIRT1 was essential for RSV to protect cells against HG through the regulation of MG- and ROS-targeting defences.
Human umbilical vein endothelial cells (HUVECs) were kept in 5.55 mM glucose (CTR) or 30.55 mM glucose (HG), and co-incubated with either RSV (5 µM) or RSV+EX527 (SIRT1 inhibitor) (5 µM+13.4 µM, respectively). Through morphological, molecular and biochemical methods, we evaluated: cell viability, apoptosis, expression of SIRT1, CAT (catalase), SOD1 (Cu/Zn-superoxide dismutase), SOD2 (mitochondrial Mn-superoxide dismutase), glyoxalase 1 (GLO1), ROS- and MG-dependent damages, and redox balance of glutathione. The involvement of mitochondria was verified by investigating the expression of SIRT3 and acetylated SOD2.
We found that HG impaired MG/ROS scavenging activities in HUVECs, and that RSV effectively rescued such impairments by up-regulating SIRT1. Interestingly, the response involved mitochondria, as seen by both changed expression of SIRT3 and altered levels of SOD2.