毛钩藤碱通过激活线粒体自噬减轻高脂诱导的H9C2细胞凋亡  

Hirsutine mitigates high-fat-induced apoptosis in H9C2 cells by activating mitophagy

作　　者：盛孙朋 陈诗佳 董希诚 丁露 马建设 金培峰[1] SHENG Sunpeng;CHEN Shijia;DONG Xicheng;DING Lu;MA Jianshe;JIN Peifeng(Department of Cardiac Surgery,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China;The First School of Medicine,School of Information and Engineering,Wenzhou Medical University,Wenzhou 325035,China;School of Pharmaceutical Science,Wenzhou Medical University,Wenzhou 325035,China;School of Basic Medical Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学附属第一医院心脏外科,浙江温州325015 [2]温州医科大学第一临床医学院(信息与工程学院),浙江温州325035 [3]温州医科大学药学院,浙江温州325035 [4]温州医科大学基础医学院,浙江温州325035

出　　处：《温州医科大学学报》2024年第10期791-797,805,共8页Journal of Wenzhou Medical University

基　　金：温州市基础性科研项目(Y2020019)。

摘　　要：目的:探讨毛钩藤碱对高脂诱导H9C2细胞线粒体自噬及凋亡的作用及影响。方法:未经任何处理的H9C2细胞作为空白对照(Control)组,榈酸刺激H9C2细胞作为高脂(PA)组,毛钩藤碱处理细胞作为治疗(PA+HS)组,Rho激酶抑制剂Y-27632处理细胞为抑制剂(PA+HS+Y-27632)组。CCK8法评估细胞生长状态;TUNEL染色检测细胞凋亡情况;TMRE染色检测线粒体膜电位(mPTP)的变化;免疫印记法检测细胞内线粒体自噬蛋白LC3 I/II以及PINK1、Parkin信号通路,检测凋亡相关蛋白Cleaved-caspase 3、caspase 3、Bax、Bcl-2表达水平变化。结果:与Control组比较,200、400、800μmol/LPA处理细胞的活力均有所下降(P<0.05);与Control组比较,PA组细胞TMRE染色荧光强度显著变弱,LC3-I/II、PINK以及Parkin表达水平显著降低(P<0.05),TUNEL染色显示PA组阳性细胞显著增加,Cleaved-caspase 3/caspase 3、Bax/Bcl-2表达水平显著增高(P<0.05);与PA组比,PA+HS组细胞TMRE染色荧光强度显著变强,LC3-I/II、PINK以及Parkin的表达水平显著增高(P<0.05),TUNEL染色显示PA+HS组阳性细胞显著降低,Cleaved-caspase 3/caspase 3、Bax/Bcl-2表达水平显著降低(P<0.05);与PA+HS组比,PA+HS+Y-27632组的TMRE染色荧光强度减弱,LC3-I/II、PINK以及Parkin表达水平显著降低(P<0.05),TUNEL染色显示PA+HS+Y-27632组阳性细胞显著升高,Cleaved-caspase 3/caspase 3、Bax/Bcl-2表达水平显著升高(P<0.05)。结论:毛钩藤碱通过激活线粒体自噬减轻高脂诱导H9C2细胞凋亡。Objective:To explore the effect of Hirsutine on mitophagy and apoptosis in H9C2 cells induced by high-fat.Methods:With untreated H9C2 cells served as the control group(Control),H9C2 cells stimulated by palmitic acid were used as the high-fat group(PA),H9C2 cells treated with Hirsutine as the treatment group(PA+HS)and H9C2 cells treated with the Rho kinase inhibitor Y-27632 as the inhibitor group(PA+HS+Y-27632).CCK8 assay was used to evaluate cell viability;TUNEL staining was used to detect cell apoptosis;TMRE staining was used to detect changes in mitochondrial membrane potential(mPTP);Western blot was used to detect the expression of mitophagy proteins LC3I/II,PINK 1 and Parkin as well as apoptosis-related proteins Cleaved-caspase 3,caspase 3,Bax,and Bcl-2.Results:Compared with the control group,the viability of cells treated with 200,400,and 800μmol/L PA decreased(P<0.05);the TMRE fluorescence intensity was significantly weaker,while the expression of LC3-I/II,PI,and Parkin was significantly lower(P<0.05);the TUNEL-positive cells increased significantly,and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 increased significantly(P<0.05)in the PA group.Compared with the PA group,the TMRE fluorescence intensity was significantly stronger;the expression of LC3-I/II,PI,and Parkin was significantly higher(P<0.05);the TUNEL-positive cells decreased significantly,and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 decreased significantly(P<0.05)in the PA+HS group.Compared with the PA+HS group,the TMRE fluorescence intensity was weaker;the expression of LC3-I/II,PI,and Parkin was significantly lower(P<0.05);the TUNEL-positive cells increased significantly,and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 increased significantly(P<0.05)in the PA+HS+Y-27632 group.Conclusion:Hirsutine alleviates apoptosis of H9C2 cells induced by high-fat through the activation of mitophagy.

关 键 词：毛钩藤碱 心肌细胞 线粒体自噬 细胞凋亡 

分 类 号：R363.2[医药卫生—病理学]