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红杉醇对高糖诱导的人脐静脉内皮细胞NOX4、eNOS表达的影响
引用本文:李先伟,郝伟,刘艳,杨人. 红杉醇对高糖诱导的人脐静脉内皮细胞NOX4、eNOS表达的影响[J]. 中国应用生理学杂志, 2014, 0(2): 147-152
作者姓名:李先伟  郝伟  刘艳  杨人
作者单位:皖南医学院药理学教研室,安徽芜湖241002
基金项目:安徽省优秀青年基金项目(2011SQRL105)
摘    要:目的:观察红杉醇(Scq)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1h后,30mmol/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(R0s)水平,比色法检测细胞-氧化氮(NO)、丙二醛(MDA)及过氧化氢(H202)水平,real-timePCR和Westernblot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1h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H202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。

关 键 词:红杉醇  人脐静脉内皮细胞  NOX4  eNOS  氧化应激

Effect of sequoyitol on expression of NOX4 and eNOS induced with glucose in human umbilical vein endothelial cells
Affiliation:LI Xian-wei, HAO Wei, LIU Yan, YANG Jie-ren (Department of Phanmcology, Wannan Medical College, Wuhu 241002, China)
Abstract: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.
Keywords:sequoyitol  human umbilical vein endothelial cells  NOX4  eNOS  oxidative stress
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