NADPH氧化酶4在高糖促进HUVECs内活性氧产生机制中的作用

The role of NADPH oxidase in intracellular ROS generation stimulated by high concentration of glucose

目的:研究在高浓度葡萄糖刺激状态下,人脐静脉内皮细胞(HUVECs)内活性氧的主要来源。方法:Ⅱ型胶原酶消化法分离培养人脐静脉内皮细胞,流式细胞术检测细胞内活性氧的生成,RT-PCR及Westernblot检测细胞内NADPH氧化酶4(NOX4)的表达。结果:高糖处理HUVECs后细胞内ROS升高、NOX4表达明显上调;NOX抑制剂DPI能够抑制NOX4表达的上调和细胞内ROS的升高。结论:高糖处理HUVECs时细胞内ROS升高主要来源于NOX4。

Objective： To study the main source of ROS generated in human umbilical vein endothelial cells （HUVECs） while stimulated by high concentration of glucose.Methods：Human umbilical vein endothelial cells was isolated by type Ⅱ collagenase, Intracellular ROS generation was measured by flow cytometry, NOX4 expression in HUVECs was detected by RT-PCR and Western blot.Results：High concentration of glucose could increase ROS generation and NOX4 expression; DPI, the specially inhibitor of NADPH oxidase, could attenuated these effects. Conclusion：NOX4 was the main source of ROS generation in HUVECs cultured with high concentration of glucose.