Lei Wang1, Huabin Su, Qiong Lu, Heping Wu. 1. Department of Basic Medicine, Hunan University of Medicine, Huaihua Hunan 418000, China wanglei800129@aliyun.com.
Abstract
OBJECTIVE: To explore the effect of high glucose on NADPH oxidase (NOX) expression and intracellular reactive oxygen species (ROS) generation in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were divided into a control group, a mannite group, a glucose group and a glucose plus diphenylene iodonium (DPI) group. Intracellular ROS was detected by flow cytometry. RNA and protein expression of NOX in HUVECs was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: 1) Compared with the control group, the intracellular ROS were significantly increased in the glucose group (P< 0.05, n=3), but there were no significant change in the glucose plus DPI group (P> 0.05, n=3); 2) Compared with the control group, the mRNA and protein expression of NOX4 in the glucose group were significantly increased (P< 0.05), but there were no significant change in the mRNA and protein expression of NOX2, p22phox, p67phox and rac (all P> 0.05); 3) there were no significant difference in the mRNA and protein expression of NOX2, NOX4, p22phox, p67phox and rac between the glucose plus DPI group and the control group (all P> 0.05). CONCLUSION: High glucose may increases intracellular ROS generation by increasing the expression of NOX4 in HUVECs, which might mediate the oxidative stress.
OBJECTIVE: To explore the effect of high glucose on NADPH oxidase (NOX) expression and intracellular reactive oxygen species (ROS) generation in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were divided into a control group, a mannite group, a glucose group and a glucose plus diphenylene iodonium (DPI) group. Intracellular ROS was detected by flow cytometry. RNA and protein expression of NOX in HUVECs was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: 1) Compared with the control group, the intracellular ROS were significantly increased in the glucose group (P< 0.05, n=3), but there were no significant change in the glucose plus DPI group (P> 0.05, n=3); 2) Compared with the control group, the mRNA and protein expression of NOX4 in the glucose group were significantly increased (P< 0.05), but there were no significant change in the mRNA and protein expression of NOX2, p22phox, p67phox and rac (all P> 0.05); 3) there were no significant difference in the mRNA and protein expression of NOX2, NOX4, p22phox, p67phox and rac between the glucose plus DPI group and the control group (all P> 0.05). CONCLUSION: High glucose may increases intracellular ROS generation by increasing the expression of NOX4 in HUVECs, which might mediate the oxidative stress.