锦葵色素葡萄糖苷和锦葵色素半乳糖苷抑制内皮细胞氧化应激作用

Effects of Malvidin Glycosides on Ang Ⅱ-Induced Oxidative Stress in Endothelial Cells

目的:研究血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)中活性氧(reactive oxygen species,ROS)、黄嘌呤氧化酶-1(xanthine oxidase-1,XO-1)、血红素氧化酶-1(heme oxygenase-1,HO-1)含量和超氧化物歧化酶(superoxide dismutase,SOD)活性的影响,确定最佳AngⅡ诱导浓度,探讨不同浓度锦葵色素葡萄糖苷(malvidin-3-glucoside,Mv-3-glc)和锦葵色素半乳糖苷(malvidin-3-galactoside,Mv-3-gal)对AngⅡ诱导的血管内皮细胞氧化应激的抑制作用机理。方法:通过体外培养HUVEC,采用酶联免疫吸附分析法分别测定HUVEC中SOD活力、XO-1和HO-1含量,并采用荧光探针方法检测细胞中ROS水平变化。结果:在HUVEC中,AngⅡ可诱导XO-1的表达,抑制HO-1的表达,AngⅡ在浓度10-2 μmol/L时对HUVEC氧化应激诱导最为显著。不同浓度的Mv-3-glc、Mv-3-gal及其混合物均可显著抑制AngⅡ诱导的内皮细胞氧化应激,通过抑制XO-1蛋白过表达,增加HO-1和SOD表达量,改善细胞抗氧化防御系统,从而减少细胞内ROS水平,达到氧化保护作用,且Mv-3-glc比Mv-3-gal抗氧化活性更强。结论:锦葵色素葡萄糖苷和锦葵色素半乳糖苷具有抑制内皮细胞氧化应激的作用。

The objective of this research was to investigate the antioxidant role in endothelial cells. The influence of angiotensin Ⅱ（Ang Ⅱ） on the expression of xanthine oxidase-1（XO-1） and heme oxygenase-1（HO-1） in human umbilical vein endothelial cells（HUVECs）, and the changes in reactive oxygen species（ROS）, superoxide dismutase（SOD）, XO-1, and HO-1 caused by two malvidin glycosides malvidin-3-glucoside（Mv-3-glc） and malvidin-3-galactoside（Mv-3-gal） were investigated. The levels of XO-1, HO-1, and SOD were evaluated by enzyme linked inmunosorbent assay（ELISA） kits. ROS level was detected under a fluorescence microplate. Compared with the control group, the expression of XO-1 protein was significantly increased and the expression of HO-1 protein was significantly decreased in the 10-2 μmol/L Ang Ⅱ-treated group. Different concentrations of Mv-3-glc and Mv-3-gal as well as their mixture greatly decreased the levels of ROS and XO-1, but increased the levels of SOD and HO-1. Glycosides improved the antioxidant capacity of malvidin to a great extent. The antioxidant activity of Mv-3-glc was more pronounced than that of Mv-3-gal. Variation in malvidin glycosides had a significant impact on Ang Ⅱ-induced oxidative stress to different extents. Therefore, malvidin glycosides can protect cells from oxidative deterioration and thus used to prevent diseases associated with oxidative stress.