穗花杉双黄酮对过氧化氢诱导血管内皮细胞氧化应激损伤的影响

Effect of amentoflavone on H_(2)O_(2)-induced oxidative stress injury of vascular endothelial cells

目的探讨穗花杉双黄酮(AF)对过氧化氢诱导血管内皮细胞氧化应激损伤的保护作用及相关机制。方法设对照组、过氧化氢(H_(2)O_(2))组、H_(2)O_(2)+AF组,H_(2)O_(2)+AF组用不同浓度(5、10μm)的AF干预H2O2诱导的血管内皮细胞,对照组不加AF处理。细胞计数试剂盒8法检测计算出AF对细胞的安全浓度,酶联免疫吸附法检测干预后细胞上清液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)各自的表达水平,Woundhealing实验观察干预后细胞迁移能力,荧光染色观察干预后细胞凋亡小体,流式细胞术检测干预后细胞凋亡情况,Westernblot检测Bcl-2、Bax、Bcl-x、Cleaved-caspase 3凋亡相关蛋白表达。结果AF干预后可以改善被H2O2诱导引起的内皮细胞迁移能力,使SOD活性水平上调,LDH表达水平下调;AF可抑制H_(2)O_(2)诱导的内皮细胞凋亡,调节Bax和Cleaved-caspase 3蛋白下调,促进Bcl-2和Bcl-x上调。结论AF可通过抑制细胞内氧化应激水平而保护过氧化氢诱导的血管内皮细胞损伤。

Objective To explore the protective effect of amentoflavone(AF)on H2O2-induced oxidative stress injury of vascular endothelial cells and the mechanism.Methods A control group,a H2O2 group,and a H_(2)O_(2)+AF group were set.The H_(2)O_(2)-induced vascular endothelial cells in the H_(2)O_(2)+AF group were intervened with 5 and 10μm AF,and those in the control group were not intervened with AF.The cell counting kit-8 method was used to detect and calculate the safe concentration of AF.The enzyme-linked immunosorbent assay was used to detect the levels of cell supernatant lactate dehydrogenase(LDH)and superoxide dismutase(SOD).The The prognostic cell migration ability was observed by Wound healing experiment.The apoptotic bodies were observed by fluorescence staining.The apoptosis after the intervention was detected by flow cytometry.The Bc1-2,Bax,Bcl-x,and Cleaved-caspase 3 were detected by Western blot.Results AF improved the cells'migration ability,increased the level of SOD activity,and down-regulated the expression level of LDH.AF depressed the cells'apoptosis,down-regulated the expression of Bax and Cleaved-caspase 3 proteins,and up-regulated Bcl-2 and Bcl-x.Conclusion AF protects the H2O2-induced oxidative stress injury of vascular endothelial cells by inhibiting the cells'oxidative stress.