成纤维细胞生长因子19改善缺氧/复氧心肌细胞H9c2损伤

目的:研究成纤维细胞生长因子19(FGF19)对缺氧/复氧心肌细胞H9c2损伤的改善作用。方法:将心肌细胞H9c2分为Control组(常规方法培养)、H/R组(细胞经缺氧/复氧处理)、NC+H/R组(细胞转染pcDNA 3.1后再经缺氧/复氧处理)和FGF19+H/R组(细胞转染pcDNA 3.1-FGF19后再经缺氧/复氧处理)。实时荧光定量PCR(RT-qPCR)和蛋白印迹(Western blot)检测各组FGF19 mRNA和蛋白表达水平,ELISA检测各组细胞培养液中乳酸脱氢酶(LDH)含量和细胞中丙二醛(MDA)、活性氧(ROS)、LDH、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)水平,流式细胞术测定各组细胞凋亡,Western blot检测各组细胞中活化的半胱天冬酶-3(C-Caspase-3)、活化的半胱天冬酶-9(C-Caspase-9)蛋白表达和胞浆中细胞色素C(Cyt C)蛋白水平,JC-1法检测各组细胞线粒体膜电位变化。结果:与H/R组和NC+H/R组比较,FGF19+H/R组心肌细胞中FGF19 mRNA和蛋白表达水平均显著升高(P均<0.01)。H/R组与NC+H/R组心肌细胞中FGF19 mRNA和蛋白表达水平比较差异无统计学意义(P>0.05)。与Control组比较,H/R组细胞培养液中LDH水平升高(P<0.01),细胞中MDA和ROS水平升高(P<0.01),SOD、CAT和GSH-Px活力降低(P<0.01),细胞凋亡率、C-Caspase-3、C-Caspase-9和Cyt C蛋白水平升高(P<0.01),线粒体膜电位降低(P<0.01)。与NC+H/R组比较,FGF19+H/R组细胞培养液中LDH水平降低(P<0.01),细胞中MDA和ROS水平降低(P<0.01),SOD、CAT和GSH-Px活力升高(P<0.01),细胞凋亡率、C-Caspase-3、C-Caspase-9和Cyt C蛋白水平降低(P<0.01),线粒体膜电位升高(P<0.01)。结论:FGF19可能通过减少细胞凋亡和氧化损伤改善缺氧/复氧心肌细胞H9c2损伤。

Fibroblast Growth Factor 19 Improves H9c2 Injury in Hypoxia/Reoxygenation Cardiomyocytes

Objective:To study the protective effect of fibroblast growth factor 19(FGF19)on hypoxic/reoxygenated cardiomyocyte H9c2 injury.Method:Cardiomyocytes H9c2 were divided into control group,H/R group,NC+H/R group and FGF19+H/R group.qRT-PCR and Western blot were used to detect the expression of FGF19 mRNA and protein in each group.The LDH content in the cell culture medium and the levels of MDA,ROS,LDC,SOD,and CAT in cells of each group were detected by ELISA.Flow cytometry was used to determine the apoptosis of each group.Western blot was used to detect the levels C-Caspase-3,C-Caspase-9 and Cyt C protein.The mitochondrial membrane potential changes of each group of cells were detected by JC-1 method.Results:Compared with H/R group and NC+H/R group,the expression of FGF19 mRNA and protein in FGF19+H/R group were significantly increased(P<0.01).There was no significant difference in the expression of FGF19 mRNA and protein between H/R group and NC+H/R group(P>0.05).Compared with the control group,the LDH level in the H/R group was increased(P<0.01),the levels of MDA and ROS in the cells were increased(P<0.01),the levels of SOD,CAT and GSH-Px were decreased(P<0.01),apoptosis rate and the levels of C-Caspase-3,C-Caspase-9 and Cyt C protein were increased(P<0.01),and mitochondrial membrane potential was decreased(P<0.01).Compared with the NC+H/R group,the LDH level in the FGF19+H/R group was decreased(P<0.01),the levels of MDA and ROS in the cells were decreased(P<0.01),the levels of SOD,CAT and GSH-Px were increased(P<0.01),apoptosis rate and the levels of C-Caspase-3,C-Caspase-9 and Cyt C protein were decreased(P<0.01),and mitochondrial membrane potential was increased(P<0.01).Conclusion:FGF19 can significantly improve H9c2 damage in hypoxic/reoxygenated cardiomyocytes by reducing apoptosis and oxidative damage.