The Effects Of P22phox On Endothelial Cell Apoptosis Induced By High-glucose And Its Mechanisms

Objectives: To study the effects of NAD (P)H oxidase subunits p22phox on endothelial ROS induction and cell apoptosis induced by high-glucose.Methods:Mophology of HUVECs was observed by inverted microscope. Expression of factorâ…§related antigen in HUVECs was investigated by immunohistochemistry. HUVECs were divided into four groups:control group,high glucose group , siRNA group and high glucose+siRNA group. Expression of p22phox and caspase-3 were investigated by western-blot. Intracellular ROS level, and cell apoptosis rate in HUVECs were detected by flow cytometry. Hochest33342 fluorescent dying was used to detect cell apoptosis. Activity of caspase 3 was detected by fluoremetry.Results :The HUVECs were detached successfully. p22phox-siRNA could induce NAD(P)H oxidase subunit p22phox gene silence.High glucose could induce p22phox expression. Compared with control group, p22phox expression ,ROS level ,cell apoptosis rate,caspase 3 activity and caspase-3 expression was significantly increased in high glucose group(P < 0.05). But p22phox expression ,ROS level ,cell apoptosis rate,caspase 3 activity and caspase-3 expression in high glucose+siRNA group significantly decreased (P < 0.05)compared with high glucose group(P < 0.05).Conclusions : Transfection with siRNA-p22phox could effectively inhibit p22phox expression in HUVECs.Inhibiting p22phox expression could decrease ROS level and cell apoptosis induced by high-glucose.