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The Effects of p22phox on Endothelial Cell Apoptosis Induced by High-glucose and Its Mechanisms

Author: ShengShunLiang
Tutor: WangZuo
School: Nanhua University
Course: Pathology and Pathophysiology
Keywords: Atherosclerosis p22phox Human Umbilical Vein Endothelial Cells Glucose Reactive Oxygen Species
CLC: R363
Type: Master's thesis
Year: 2009
Downloads: 17
Quote: 0
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Abstract


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 factorⅧrelated 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.

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