达格列净对D-半乳糖诱导心肌细胞衰老和氧化应激的影响

Effects of Dapagliflozin on Myocardial Cell Senescence and Oxidative Stress Induced by D-galactose

目的探讨达格列净对D-半乳糖诱导心肌细胞衰老和氧化应激的影响。方法体外培养H9C2心肌细胞,使用D-半乳糖构建心肌细胞衰老模型。将细胞分为正常对照组(采用正常细胞培养液培养)、衰老模型组(采用10 g/L D-半乳糖培养细胞72 h)以及2、10、50μmol/L达格列净组(分别采用2、10、50μmol/L达格列净与10 g/L D-半乳糖共同培养细胞72 h)。通过衰老相关β-半乳糖苷酶染色评估达格列净对心肌细胞衰老的影响;通过流式细胞术检测细胞内活性氧(ROS)含量、黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)活性、Western blotting检测p21、血红素加氧酶-1(HO-1)、醌氧化还原酶1(NQO1)蛋白表达变化。结果与正常对照组比较,衰老模型组的衰老相关β-半乳糖苷酶染色阳性细胞比例、ROS含量、p21蛋白表达升高,SOD活性、HO-1和NQO1蛋白表达降低(P<0.05,P<0.01);与衰老模型组比较,10、50μmol/L达格列净组衰老相关β-半乳糖苷酶染色阳性细胞比例、p21蛋白表达降低(P<0.05,P<0.01);10μmol/L达格列净组细胞中ROS含量降低,SOD活性、HO-1和NQO1蛋白表达升高(P<0.05)。结论达格列净可以延缓D-半乳糖诱导H9C2心肌细胞衰老,改善细胞氧化应激。

Objective To investigate the effects of Dapagliflozin on cell senescence and oxidative stress of cardiomyocytes induced by D-galactose.Methods H9C2 cardiomyocytes were cultured in vitro and D-galactose was used to construct cardiomyocyte senescence model.The cells were divided into normal control group(cultured with normal cell culture medium),senescence model group(cultured with 10 g/L D-galactose for 72 h)and 2,10,and 50μmol/L Dapagliflozin groups(cultured with 2,10,and 50μmol/L Dapagliflozin and 10 g/L D-galactose,respectively for 72 h).The effects of Dapagliflozin on myocardial cell senescence were evaluated by senescence-relatedβ-galactosidase staining.Intracellular reactive oxygen species(ROS)content was detected by flow cytometry,superoxide dismutase(SOD)activity was detected by xanthine oxidase,and protein expression changes of p21,heme oxygenase-1(HO-1)and quinone oxidoreductase 1(NQO1)were detected by Western blotting.Results Compared with normal control group,the proportion of senescence-relatedβ-galactosidase staining positive cells,ROS content and p21 protein expression were increased in senescence model group,while SOD activity,HO-1 and NQO1 protein expressions were decreased(P<0.05,P<0.01).Compared with senescence model group,the proportion of senescence-relatedβ-galactosidase positive cells and the expression of p21 protein in 10 and 50μmol/L Dapagliflozin groups were decreased(P<0.05,P<0.01).ROS content was decreased in 10μmol/L Dapagliflozin group,while SOD activity,HO-1 and NQO1 protein expression were increased(P<0.05).Conclusion Dapagliflozin can delay D-galactose-induced senescence of H9C2 cardiomyocytes and improve cellular oxidative stress.