Irisin通过Sirt1/UCP2拮抗二氯化钴诱导的心肌损伤作用  

Irisin inhibits myocardial injury induced by cobalt dichloride through regulating Sirt1/UCP2 pathway

作　　者：李如利 黄韵晓 韩彬 刘福[1,2] LI Ru-li;HUANG Yun-xiao;HAN Bin;LIU Fu(North Sichuan Medical College,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan 637000,China;Dept of Pharmacy,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan 637000,China;Dept of Clinic Laboratory,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan 637000,China)

机构地区：[1]川北医学院,川北医学院附属医院,四川南充637000 [2]川北医学院附属医院药剂科,四川南充637000 [3]川北医学院附属医院检验科,四川南充637000

出　　处：《中国药理学通报》2023年第6期1061-1066,共6页Chinese Pharmacological Bulletin

基　　金：基金资助:国家自然科学基金资助项目(No 81900276);四川省卫生健康委科研普及应用项目(No 20PJ146);川北医学院附属医院重点项目(No 2019-9)。

摘　　要：目的探讨肌肉因子Irisin调节胞内保护蛋白Sirt1和线粒体解偶联蛋白2(uncoupling protein 2,UCP2)在心肌缺氧过程中发挥的作用及具体机制。方法以CoCl_(2)作用H9c2心肌细胞24 h,构建心肌细胞体外缺氧模型。实验分为6组:control组、Irisin 10 nmol·L^(-1)组、Irisin 20 nmol·L^(-1)组、CoCl_(2)模型组、CoCl_(2)+Irisin 10 nmol·L^(-1)治疗组、CoCl_(2)+Irisin 20 nmol·L^(-1)治疗组。CCK-8法检测细胞存活率,Flow cytometry测定细胞凋亡和线粒体膜电位,探针法指示细胞内活性氧(reactive oxygen species,ROS)高低水平,Western blot法检测细胞内Sirt1、UCP2、Bcl-2、Bax、cleaved-caspase3的表达水平。结果心肌细胞H9c2缺氧模型组细胞活力下降、细胞凋亡增多、ROS增加、线粒体膜电位明显下降,同时抗凋亡蛋白Bcl-2、Sirt1和UCP2表达下调,凋亡相关蛋白Bax、cleaved-caspase3表达上调;与模型组相比,Irisin预保护治疗组发挥保护作用呈浓度依赖性,表现为能明显抑制细胞凋亡,改善细胞活力,同时抑制ROS水平,恢复线粒体膜电位,上调保护性蛋白Sirt1、UCP2、Bcl-2的表达,下调介导凋亡的Bax、cleaved caspase-3的表达。结论Irisin通过激活Sirt1/UCP2,进而抑制CoCl_(2)诱导的线粒体膜电位下降,改善氧化应激介导的心肌细胞凋亡。Aim To investigate the role and specific mechanisms of muscle factor Irisin in regulating the intracellular protective protein Sirt1 and mitochondrial uncoupling protein 2(UCP2)during myocardial hypoxia.Methods H9c2 cells were treated with CoCl2 for 24 hours to construct an in vitro hypoxia model of myocardial cells.Six groups were divided in this experiment:control group(control),Irisin group(10 nmol·L^(-1)),Irisin group(20 nmol·L^(-1)),model group(CoCl_(2)),irisin(10 nmol·L^(-1))+CoCl_(2) group and Irisin(20 nmol·L^(-1))+CoCl_(2) group.CCK-8 method was used to detect the cell survival rate.Flow cytometry was used to detect apoptosis and mitochondrial membrane potential.DCFH-DA probe method was used to indicate the level of reactive oxygen species(ROS)in cells and Western blot was used to detect the expression of Sirt1,UCP2,Bcl-2,Bax and cleaved-caspase3.Results In the H9c2 hypoxia model group,the cell viability decreased,apoptosis increased,ROS increased,and mitochondrial membrane potential decreased significantly.At the same time,the expression of anti-apoptotic proteins Bcl-2,Sirt1,and UCP2 was down regulated,and the expression of apoptosis related proteins Bax and cleaved-caspase3 was up regulated.Compared with the model group,the protective effect of irisin was concentration dependent,which was manifested by significantly inhibiting cell apoptosis,improving cell viability,inhibiting ROS level,restoring mitochondrial membrane potential,up regulating the expression of protective proteins Sirt1,UCP2,Bcl-2,and down regulating the expression of Bax and cleaved-caspase-3.Conclusion Irisin can inhibit the decrease of mitochondrial membrane potential induced by CoCl_(2) by activating Sirt1/UCP2,and improve cardiomyocyte apoptosis mediated by oxidative stress.

关 键 词：Irisin 氧化应激 凋亡 线粒体膜电位 SIRT1 UCP2 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学] R364.12R349.1R542.22