Study on time-effects of Tanshinone ⅡA on oxidative stress EA.Hy926 cell model’s autophagy
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摘要: 目的:研究丹参酮ⅡA对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(EA.Hy926)氧化应激模型自噬效应的影响及时效关系。方法:①采用MTT法分别观察不同浓度ox-LDL(模型组)及丹参酮ⅡA(丹参酮组)对EA.Hy926细胞增殖的影响,明确ox-LDL及丹参酮ⅡA作用于EA.Hy926细胞的浓度;②采用荧光免疫法细胞技术及免疫印迹技术检测细胞内LC3蛋白不同时间点6 h、12 h、24 h表达的变化。结果:100 mg/L ox-LDL作用于EA.Hy926细胞24 h造成明显氧化应激损伤,丹参酮ⅡA能减轻此损伤。LC3/DAPI复染发现丹参酮ⅡA组LC3自噬小体在6 h、12 h、24 h形成较模型组增多,在12 h表达最多。免疫印迹技术发现LC3A/B蛋白表达水平在12 h及24 h时明显增高,但在12 h时自噬效应蛋白LC3A/B表达最为明显。结论:丹参酮ⅡA上调EA.Hy926氧化应激细胞模型的自噬,保护细胞氧化应激损伤,并存在时效关系。Abstract: Objective:To study the time-effect of Tanshinone ⅡA on autophagy of oxidative stress human umbilical vein endothelial cells (EA.Hy926) model induced by ox-LDL. Method:①MTT method was used to observe the effect of different concentration of ox-LDL and tanshinoneⅡA on EA.Hy926 cells proliferation. We figured out these concentrations of ox-LDL and tanshinoneⅡA. ②Fluorescence and Western blot were observed the expressions of autophagy marker protein LC3 at 6 h, 12 h and 24 h. Result: One hundred mg/L ox-LDL caused significantly oxidative stress damage in human umbilical vein endothelial cells for 24 h, but tanshinoneⅡA reduced the oxidative injury induced by ox-LDL. LC3/DAPI staining found that in the tanshinoneⅡA group LC3 autophagy bodies formation were increased at 6-24 h, peaked at 12 h, when compared with the model group. Western blot found that autophagy marker protein LC3A/B expression was increased obviously at 12 h in the tanshinoneⅡA group. Conclusion:Tanshinone ⅡA affects the autophagy of oxidative stress EA.Hy926 cells modle with time-effect. Tanshinone ⅡA protects EA. Hy926 cells from oxidative damage by autophagy upregulation.
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Key words:
- tanshinoneⅡA /
- endothelial cell /
- atherosclerosis /
- oxidative stress /
- autophagy
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