丹参酮ⅡA逆转高糖诱导一氧化氮生成减少及机制研究

摘要

Objective: To investigate the effect of TanshinoneⅡA on nitric oxide (NO) production in human umbilical vein endothelial cells atfer high glucose treatment.Methods:HUVECs were randomly divided into ifve groups: control, mannitol, high glucose, high glucose + vehicle, high glucose + tanshinoneⅡA. NO production was determined by 3-Amino, 4-aminomethyl-2', 7'-difluorescein, diacetate (DAF-FM DA) with a laser scanning confocal microscope. cGMP concentration and total nitrite production were examined by ELISA assay and Griess reagent, respectively. Endothelial nitric oxide synthase (eNOS) expression was detected by qPCR and western blotting, respectively.Results: TanshinoneⅡA signiifcantly restored high glucose-induced decrease of NO production, cGMP concentration and total nitrite production. Additionally, TanshinoneⅡA inhibited high glucose-induced decrease of eNOS protein expression, whereas produced no effects on eNOS mRNA expression. Conclusion:TanshinoneⅡA restores high glucose-induced decrease of nitric oxide production via enhancing eNOS protein expression.%目的：观察丹参酮ⅡA对高糖引起内皮细胞中一氧化氮生成量减少的影响，并探讨相关的作用机制。方法：将内皮细胞随机分为空白对照组，等渗对照组，高糖处理组，溶剂对照组，丹参酮ⅡA组。一氧化氮特异性探针检测法检测内皮细胞一氧化氮含量。分别采用酶联免疫吸附(ELISA)和Griess法检测环磷酸鸟苷(cGMP)和总硝基化合物的含量。免疫印迹法和实时荧光定量聚合酶链反应(qPCR)分别检测内皮型一氧化氮合酶(endothelial nitric oxide synthase，eNOS)蛋白和mRNA水平的变化。结果：丹参酮ⅡA可抑制高糖诱导的人脐静脉内皮细胞中一氧化氮生成量(P<0.05)， cGMP含量以及总硝基化合物生成的减少(P<0.05)，并能逆转eNOS蛋白表达的降低(P<0.05)。结论：丹参酮ⅡA通过上调eNOS蛋白表达抑制一氧化氮生成的减少。