Protective Effects Of GSH On Endothelial Cell Oxidative Damage Cultured In MODS Serum

ObjectiveTo investigate the effects of oxidative damage in human umbilical vein endothelial cells (HUVEC) cultured in the patient serum with multiple organ dysfunction syndrome (MODS) and the protective effects of GSH on the endothelial dysfunction.MethodsTo select MODS patients and healthy control (n=20), HUVEC were cultured in the serum of patients with MODS to establish the injury model. At the same time healthy serum and fetal bovine serum groups were used as control. Cells were divided into normal control group(group A), healthy serum group(group B), MODS serum injury group(serum concentration respectively was groupCl:5%, groupC2:10%, group C3:20%) and GSH intervention group(GSH concentration separately was group D1:0.064mmol/L, D2:0.32mmol/L, D3:1.6mmol/L, D4:8.0mmol/L, D5:40mmol/L).The proliferative activity of HUVEC was assessed by blue tetrazolium bromide (MTT).The cell morphology were observed by inverted microscope in group B and Group C2. The supernatant concentrations of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) were measured in group B, group C2and group D3after GSH intervention24,48and72h respectively, while intracellular reactive oxygen species (ROS) level was assessed by flow cytometry.Result1. There were no significant difference in cell survival rate between group B and group A (P>0.05). Cell survival rate of group C1, group C2and group C3was lower than in group B(P<0.05), while cell survival rate of group D2, group D3and group D4was increased higher than group C2(P<0,05).2. In group C2, The characteristic morphological changes of oxidative injury were observed in HUVEC under transmission microscope. Cells became bigger and round, the boundaries became blurred, Cell body fractured, and part of the cells fall off the bottom of the culture bottle.3. The levels of SOD, CAT, GSH-Px and T-AOC in group C2were decreased significantly, however, increased in medicine intervention after24,48and72h, especially marked in drug intervention after72hours. 4. The ROS level in group C2was obviously increased compared with group B, while evidently decreased in medicine intervention after24h,48h, and72h, specially obvious in drug intervention after72hours.ConclusionHUVEC was vulnerable to MODS patient serum, exogenous GSH can effectively protect the vascular endothelial cells from undergoing oxidative damage through increasing the level of SOD, CAT, GSH-Px and T-AOC, while decreasing the level of intracellular ROS.