CTRP3通过激活SIRT1/FOXO3a途径减轻OGD/R诱导的心肌细胞损伤

CTRP3 alleviates OGD/R-induced myocardial cell injury by activating the SIRT1/FoxO3a pathway

目的:探究CTRP3在糖氧剥夺/再灌注(OGD/R)诱导的心肌细胞损伤中的作用及其可能的作用机制。方法:体外培养大鼠心肌细胞H9c2,OGD/R诱导H9c2细胞损伤。qRT-PCR法检测细胞CTRP3、SIRT1和FOXO3a mRNA表达;Western blot检测细胞CTRP3、SIRT1、FOXO3a、Bax、Bcl-2和cleaved caspase-3蛋白表达,CCK-8检测细胞存活,流式细胞术检测细胞凋亡,检测细胞CK-MB、cTnI、SOD和MDA活性、GSH-Px含量及ROS的产生。结果:与Control组相比,OGD/R组细胞中CTRP3、SIRT1和FOXO3a mRNA以及蛋白表达,细胞存活率、Bcl-2蛋白表达、SOD和GSH-Px活性明显降低,细胞凋亡率、Bax和cleaved caspase-3蛋白表达、CK-MB、cTnI和MDA含量、ROS荧光强度明显升高(P<0.05)。OGD/R组与oe-NC组细胞各指标比较,差异无统计学意义(P>0.05)。与oe-NC组相比,oe-CTRP3组细胞中CTRP3、SIRT1和FOXO3a mRNA以及蛋白表达,细胞存活率、Bcl-2蛋白表达、SOD和GSH-Px活性明显升高(P<0.05),细胞凋亡率、Bax和cleaved caspase-3蛋白表达、CKMB、cTnI和MDA含量、ROS荧光强度明显降低(P<0.05)。与oe-CTRP3组相比,oe-CTRP3+EX527组细胞中SIRT1、FOXO3a和Bcl-2蛋白表达、细胞存活率、SOD和GSH-Px活性明显降低,细胞凋亡率、Bax和cleaved caspase-3蛋白表达、CK-MB、cTnI和MDA含量、ROS荧光强度明显升高(P<0.05)。与oe-CTRP3+EX527组相比,oe-CTRP3+EX527+oe-FOXO3a组细胞中FOXO3a和Bcl-2蛋白表达,细胞存活率、SOD和GSH-Px活性明显升高,细胞凋亡率、Bax和cleaved caspase-3蛋白表达、CKMB、cTnI和MDA含量、ROS荧光强度明显降低(P<0.05)。结论:CTRP3通过激活SIRT1/FOXO3a途径促进OGD/R诱导的心肌细胞存活,抑制细胞凋亡和氧化应激。

Objective:To explore the role of CTRP3 in cardiomyocyte injury induced by oxygen glucose deprivation/reperfusion(OGD/R)and its possible mechanism.Methods:Rat cardiomyocytes H9c2 were cultured in vitro,and OGD/R induced H9c2 cell injury.qRT-PCR was used to detect the expression of CTRP3,SIRT1 and FOXO3a mRNA in cells.Western blot was used to detect the expression of CTRP3,SIRT1,FOXO3a,Bax,Bcl-2 and cleaved caspase-3 protein in cells.CCK-8 was used to detect cell survival.Flow cytometry was used to detect cell apoptosis.The kit was used to detect the CK-MB,cTnI,SOD and MDA activity,and the GSH-Px and ROS production of cells.Results:Compared with the control group,the expressions of CTRP3,SIRT1 and FOXO3a mRNA and protein,the cell survival rate,Bcl-2 protein expression,and SOD and GSHPx activities were significantly reduced(P<0.05),while the apoptosis rate,the expression of Bax and cleaved caspase-3 protein,the contents of CK-MB,cTnI and MDA,and ROS fluorescence intensity were significantly increased in OGD/Rgroup(P<0.05).There were no significant differences in cell indicators in the oe-NC group and OGD/R group(P>0.05).Compared with the oe-NC group,the CTRP3,SIRT1 and FOXO3a mRNA and protein expressions,cell survival rate,Bcl-2 protein expression,and SOD and GSH-Px activities were significantly increased,whereas the apoptosis rate,the expression of Bax and cleaved caspase-3 protein,the contents of CK-MB,cTnI and MDA,and ROS fluorescence intensity were significantly reduced in the oe-CTRP3 group(P<0.05).Compared with the oe-CTRP3 group,the SIRT1,FOXO3a and Bcl-2 protein expression,cell survival rate,SOD and GSH-Px activities in the oe-CTRP3+EX527 group were significantly reduced,while the apoptosis rate,the expression of Bax and cleaved caspase-3 protein,the contents of CK-MB,cTnI and MDA,and ROS fluorescence intensity were increased significantly(P<0.05).Compared with the oe-CTRP3+EX527 group,the FOXO3a and Bcl-2 protein expressions,the cell survival rate and SOD and GSH-Px activities were significantly increased,while the apoptosis rate,the expressions of Bax and cleaved caspase-3 protein,the contents of CK-MB,cTnI and MDA,and ROS fluorescence intensity were significantly reduced in the oe-CTRP3+EX527+oe-FOXO3a group(P<0.05).Conclusion:CTRP3 promotes the survival of cardiomyocytes induced by OGD/R and inhibits apoptosis and oxidative stress by activating the SIRT1/FOXO3a pathway.