摘要
目的:研究羟基红花黄色素A(hydroxy-safflor yellow A,HSYA)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导人脐静脉内皮细胞(HUVECs)凋亡的保护作用及机制。方法:体外培养第4～6代HUVECs,分为正常对照组、AngⅡ损伤组(1μmoL/L)及HSYA低(10μmoL/L)、中(30μmoL/L)、高(100μmoL/L)浓度预处理3 h加AngⅡ损伤组。用MTT检测HUVECs活力,激光共聚焦显微镜荧光染色法检测细胞内活性氧(ROS),试剂盒检测细胞色素c氧化酶活性,流式细胞仪分析细胞凋亡率,western blot检测Caspase-3的表达。结果:与空白对照组比较,AngⅡ损伤模型组细胞活力显著降低、ROS生成显著增加、细胞色素c活性显著增强、细胞凋亡率显著升高、Caspase-3表达显著增加(P<0.01),提示AngⅡ可诱导HUVECs凋亡。HSYA能使模型细胞增殖活力显著增加、ROS生成显著减少、细胞色素c活性明显降低、细胞凋亡率和Caspase-3表达显著降低(P<0.05或P<0.01),提示HYSA可抑制AngⅡ引起的内皮细胞凋亡。结论:HSYA可以抑制AngⅡ诱导的内皮细胞凋亡,抑制ROS生成保护细胞线粒体可能是其机制之一。
Objective：To investigate the protective effect of hydroxy-safflor yellow A（HSYA） on the apoptosis of human umbilical vein endothelial eell（HUVECs）induced by angiotcnsin Ⅱ （Ang Ⅱ ）in vitro and explore its mechanism. Methods：The HUVECs was subeultured in vitro and used for experiment that divided into five groups as follows：control group, Ang Ⅱ -injured group （ 1 μmoL/L）, low-dosage of HSYA group（ 10 μmoL/L） ,mid-dosage of HSYA group（30μmoL/L） and high-dosage of HYSA group（ 100 μmoL/L）. MTT was used to determine the HUVECs viability. Reactive oxygen species （ROS）were measured with laser scanning confocal microscopy（LSCM） ,Cytochrome C oxidase activity was detected by BCA method. Apoptosis rate of the HUVECs was analyzed by flow cytometry. The expression of apoptosis-related protein caspase-3 was measured by western blot. Results ： Compared with control group, Ang Ⅱ could increase the level of ROS,inhibit cytochrome activity and enhance caspase 3 expression in HUVECs, as a result, enhance apoptosis of HUVECs. HSYA could significantly reduce the result induced by AngⅡ in dose-dependent manner（P 〈0. 05 or P 〈0. 01 ）. Conclusion：HSYA can eliminate the effect of Ang Ⅱ and its mechanism may be related to inhibiting ROS producing, keeping mitochondrial structure and function and inhibiting apoptosis.
出处
《中药材》
CAS
CSCD
北大核心
2013年第7期1128-1131,共4页
Journal of Chinese Medicinal Materials
