RAB7/mTOR/TFEB信号通路在心肌细胞线粒体自噬中的作用及机制研究
The Mechanism of RAB7/mTOR/TFEB Signaling Pathway in Mitochondrial Autophagy of Cardio-myocytes
兰冰 1郭晓平 1褚红蔚 1韩文忠1
作者信息
- 1. 河北省沧州中西医结合医院,沧州 061001
- 折叠
摘要
目的:分析RAB7/mTOR/TFEB信号通路在心肌细胞线粒体自噬中的作用及其机制.方法:建立并培养稳定转染的RAB7过表达和阴性对照腺病毒,分别命名为对照组和过表达RAB7组.流式细胞术检测细胞ROS水平和细胞凋亡率,溶酶体和线粒体的相互作用通过共定位的荧光成像显示,应用蛋白印迹测定RAB7、mTOR、TFEB、LC3B-Ⅱ、CaMKIIδ和P62表达水平.结果:与对照组相比,过表达RAB7组mTOR表达升高,TFEB表达降低,ROS水平升高,细胞凋亡率增加,溶酶体和线粒体之间的相互作用显著增加.线粒体自噬相关的LC3B-Ⅱ、CaMKII δ和P62蛋白表达升高(P＜0.01).结论:过表达RAB7可以通过mTOR/TFEB信号通路,增加ROS释放、细胞凋亡和线粒体自噬蛋白表达而促进心肌细胞线粒体自噬激活.
Abstract
Objective:To analyze the role and mechanism of RAB7/mTOR/TFEB signaling pathway in mito-chondrial autophagy of cardiomyocytes.Method:Stable transfected RAB7 overexpression and negative control ade-novirus were established and cultured,divided into control group and RAB7 overexpression group.The ROS levels and apoptosis rate were detected by flow cytometry.The interaction between lysosome and mitochondria was re-vealed by co-localized fluorescence imaging.The expressions of RAB7,mTOR,TFEB,LC3B-Ⅱ,CaMKIIδ and P62 were determined by western blots.Results:Compared with the control group,the western blot results showed that the expression of mTOR was increased and the expression of TFEB was decreased in the RAB7 overexpression group.Moreover,the level of ROS and the rate of apoptosis were increased in the RAB7 overexpression group.Flu-orescence imaging of co-localization of lysosomes and mitochondria revealed that the interaction between lysosomes and mitochondria was significantly increased in the RAB7 overexpression group.Compared with the control group,the expression of mitophagy-related proteins LC3B-Ⅱ,CaMKⅡδ and P62 in the RAB7 overexpression group was in-creased.Conclusion:Overexpression of RAB7 can promote the activation of cardiomyocytes mitophagy by increasing ROS release,apoptosis and mitophagy protein expression through the mTOR/TFEB signaling pathway.
关键词
RAB7/mTOR/TFEB/心肌细胞/线粒体/自噬Key words
RAB7/mTOR/TFEB/Cardiomyocytes/Mitochondria/Autophagy引用本文复制引用
出版年
2024
NETL
NSTL
万方数据