红杉醇对高糖诱导的人脐静脉内皮细胞NOX4、eNOS表达的影响

Effect of sequoyitol on expression of NOX4 and eNOS induced with glucose in human umbilical vein endothelial cells

目的:观察红杉醇(Seq)对高糖诱导的人脐静脉内皮细胞(HUVECs)损伤的保护作用及机制。方法:原代培养HUVECs,红杉醇(0.1,1,10μmol/L)预处理1 h后,30 mmol/L葡萄糖诱导内皮细胞损伤。5-溴脱氧尿嘧啶核苷(BrdU)掺入法检测细胞增殖,流式细胞术检测细胞周期,2′7′-二乙酰二氯荧光素(DCFH-DA)免疫荧光法检测细胞内活性氧簇(ROS)水平,比色法检测细胞一氧化氮(NO)、丙二醛(MDA)及过氧化氢(H2O2)水平,real-time PCR和Western blot检测细胞内皮型一氧化氮合酶(eNOS)及NADPH氧化酶4(NOX4)mRNA和蛋白表达。结果:Seq预处理1 h后能明显减轻高糖诱导的血管内皮细胞损伤,促进细胞增殖,降低胞内NOX4的表达及ROS、MDA及H2O2水平,上调eNOS的表达及NO水平。结论:Seq对高糖诱导的内皮细胞损伤具有一定的保护作用,其机制可能与其抗氧化、上调eNOS的表达有关。

Objective： To investigate the protective effect and mechanism of sequoyitol（Sep） on high glucose-induced human umbilical vein endothelial cells （HUVECs） injury. Metlmds： I-IUVECs were cultured with high glucose （30 mmol/L） in the presence or absence of se quoyitol （0.1, 1 and 10 μmol/L） for 24 h. Cell proliferation was measured by BrdU marking and cell cycle was detected by flow cytometry. 2＇, 7＇-dichlomfluorescein diacetate was used to evaluate intracellular reactive oxygen species （ROS） levels. The NO, malonydialdehyde （MDA） and H202 levels were determined by colorimetric method according to the manufacturer＇s instructions. The expression of endothelial ni- tric oxide synthase（eNOS） and NADPH oxidase 4（NOX4） were measured by real-time PCR and Western blot. Results：In the present study, we found that sequoyitol pretreatment for 1 h significantly decreased cell injury, promoted cell proliferation. Meanwhile sequoyitol significantly down-regulated NOX4 expression and decreased the level of ROS, MDA and H202, and obviously increased NO levels and up-regulated eNOS expression. Com：lt^on： Sequoyitol alleviates high glucose-induced cell injuries in HUVECs via inhibiting oxidative stress and up-regulating eNOS expression.