摘要
目的基于自噬小体形成信号通路探讨丹参酮ⅡA对ox-LDL诱导内皮细胞氧化应激损伤的保护作用及机制。方法体外培养EA.hy926细胞,随机分为正常组、模型组、丹参酮ⅡA组、丹参酮ⅡA+模型组、3-MA组、模型+3-MA组、丹参酮ⅡA+模型+3-MA组。比色法检测各组细胞氧化应激损伤丙二醛(MDA)含量及超氧化物歧化酶(SOD)活力,Western blot技术检测细胞自噬小体形成信号通路相关蛋白表达情况。结果与正常组比较,模型组MDA含量增高(P<0.01),SOD活力降低(P<0.01),LC3-Ⅰ/LC3-Ⅱ蛋白含量增多(P<0.01);3-MA组LC3-Ⅰ/LC3-Ⅱ蛋白含量无明显变化(P>0.05)。与模型组比较,丹参酮ⅡA+模型组细胞中MDA含量降低(P<0.01),SOD活力增高(P<0.01),LC3-Ⅰ/LC3-Ⅱ蛋白含量增多(P<0.01);模型+3-MA组MDA含量增高(P<0.01),SOD活力降低(P<0.01),LC3-Ⅰ/LC3-Ⅱ蛋白含量减少(P<0.01)。与丹参酮ⅡA+模型组比较,丹参酮ⅡA+模型+3-MA组MDA含量增高(P<0.01),SOD活力降低(P<0.01),LC3-Ⅰ/LC3-Ⅱ蛋白含量减少(P<0.01)。与正常组比较,模型组Atg3、Atg4b、Atg7蛋白表达水平明显增高(P<0.05,P<0.01);与模型组比较,丹参酮ⅡA+模型组Atg3、Atg7、Atg5-Atg12蛋白表达水平明显增高(P<0.05,P<0.01);与丹参酮ⅡA+模型组比较,丹参酮ⅡA+模型+3-MA组Atg3、Atg7、Atg5-Atg12蛋白表达水平均明显降低(P<0.05,P<0.01)。结论丹参酮ⅡA可能通过调节EA.hy926细胞自噬小体形成信号通路即Atg12-Atg5通路和LC3-PE通路相关蛋白,发挥其保护EA.hy926细胞抗氧化应激损伤的生物学活性,进而防治动脉粥样硬化的发生发展。
Aim Based on the signaling pathway of autophagosome formation to discuss the protective effect and mechanism of Tanshinone Ⅱ A on endothelial cell oxidative stress injury. Methods Culturing EA. hy926 cells in vitro,then randomly dividing these cells into normal group,model group,tanshinone ⅡA group,tanshinone ⅡA and model group,3-MA group,model and 3-MA group,tanshinone ⅡA,model and 3-MA group. Using colorimetric method to test the cells＇ MDA content and SOD activity of oxidative stress injury. Western blot detecting the expressions of cells＇ autophagosome formation signaling pathway related protein. Results Compared with normal group,the MDA content is increased（ P〈0.01）,the SOD activity is decreased（ P〈0.01）,and the content of LC3-Ⅰ/LC3-Ⅱ protein is increased（ P〈0.01） in model group. There is no obvious change of LC3-Ⅰ/LC3-Ⅱ protein content（ P〈0.05） in 3-MA group compared with normal group. Compared with model group,the MDA content is decreased（ P〈0.01）,the SOD activity is increased（ P〈0.01） and the content of LC3-Ⅰ/LC3-Ⅱ protein is increased（ P〈0.01） in tanshinone ⅡA and model group.The MDA content is increased（ P〈0.01）,the SOD activity is decreased（ P〈0.01）,and the content of LC3-Ⅰ/LC3-Ⅱprotein is decreased（ P〈0. 01） in model and 3-MA group compared with model group. Compared with tanshinone ⅡA and model group,the MDA content is increased（ P〈0.01）,the SOD activity is decreased（ P〈0.01）,and the content of LC3-Ⅰ/LC3-Ⅱ protein is decreased（ P〈0. 01） in tanshinone Ⅱ A,model and 3-MA group. Compared with normal group,the expression of Atg3,Atg4 b,Atg7 are increased significantly in model group（ P〈0.05 or P〈0.01）. Compared with model group,the expression of Atg3,Atg7,Atg5-Atg12 are increased significantly in tanshinone Ⅱ A and model group（ P〈0.05 or P〈0. 01）. Compared with tanshinone Ⅱ A and model group,the expression of Atg3,Atg7,Atg5-Atg12 are decreased significantly in tanshinone ⅡA,model and 3-MA group（ P〈0. 05 or P〈0. 01）. Conclusion Tanshinone ⅡA may regulate the proteins of signaling pathway of autophagosome formation as well as the Atg12-Atg5 pathway and LC3-PE pathway of EA. hy926 cell to play its protective biological activities for EA. hy926 cells against oxidative stress damage,then prevent and treat the development of As.
出处
《中国动脉硬化杂志》
CAS
北大核心
2017年第3期244-249,共6页
Chinese Journal of Arteriosclerosis
基金
辽宁省自然科学基金项目(2015020394)
辽宁省高等学校优秀人才支持计划(LR2015041)
辽宁中医药大学2015年度大学生创新创业训练计划项目(201510162000066)
关键词
丹参酮ⅡA
内皮细胞
动脉粥样硬化
氧化应激
自噬小体
Tanshinone ⅡA
Endothelial cell
Atherosclerosis
Oxidative stress
Autophagosome formation
