Role of NgR mediated oxidative stress in glucose induced RGC apoptosis

Abstract

AIM: To investigate the effect of NgR mediated oxidative stress on the apoptosis of retinal ganglion cells(RGCs)induced by glucose. METHODS: RGC-5 cell were divided into 3 groups: control group(DMEM high glucose medium+10% fetal calf serum), high glucose group(DMEM high glucose medium+10% fetal calf serum+30mmol/L glucose),NEP1-40 group(DMEM high glucose medium+10% fetal calf serum+30mmol/L glucose+1μmol/L NEP1-40). Detections were performed after 3d culture: the state of cell growth was observed by microscopy. Cell viability was detected by CCK-8 kit. The apoptosis rate of RGC cells was detected by flow cytometry(FCM). The intensity of ROS of the cells were detected by fluorescence microscopy. Intracellular MDA levels and SOD activity were measured by related kits. Western blot was used to detect the expressions of Bcl-2 and Bax proteins. RESULTS: Compared with control group, high glucose group had a poor state and cell viability decreased, cell apoptosis rate significantly increased, ROS and MDA levels were significantly enhanced, SOD activity decreased, and the expression of anti-apoptotic protein Bcl-2 was decreased and the expression of pro apoptotic protein Bax was up-regulated. Compared with glucose group, after NgR expression was inhibited by NEP1-40, the oxidative stress reaction was reduced, Bcl-2/Bax was increased, the cell status was improved, the cell viability was increased, and the apoptosis rate was decreased in the NEP1-40 group(PCONCLUSION: High concentration of glucose can induce apoptosis of RGC-5 cells by NgR mediated oxidative stress reaction.

Keywords

• retinal ganglion cells
• NgR
• oxidative stress
• apoptosis