黄杞苷减轻H9c2心肌细胞的缺氧再复氧损伤

Engeletin Alleviates Hypoxia Reoxygenation Injury in H9c2 Cells

目的探讨黄杞苷对H9c2心肌细胞缺氧再复氧损伤的影响及作用机制。方法构建H9c2心肌细胞缺氧再复氧模型,将H9c2细胞随机分为常氧+溶剂组、常氧+黄杞苷组、缺氧再复氧+溶剂组和缺氧再复氧+黄杞苷组。实时荧光定量聚合酶链反应检测相关抗氧化酶(过氧化物歧化酶2、谷胱甘肽过氧化物酶1、过氧化氢酶)的mRNA水平;试剂盒检测超氧化物歧化酶、心肌损伤标志物、丙二醛以及胱天蛋白酶-3(caspase-3)水平;免疫组织化学染色检测细胞氧化应激水平;原位末端转移酶标记法染色检测细胞凋亡水平;Western blot检测相关蛋白核转录因子红系2相关因子2(Nrf2)、血红素加氧酶1(HO-1)表达水平。结果常氧+溶剂组与常氧+黄杞苷组心肌损伤标志物、氧化应激、细胞凋亡等指标及Nrf2、HO-1蛋白表达比较,无统计学差异(P>0.05);与常氧+溶剂组比较,缺氧再复氧+溶剂组心肌损伤标志物、活性氧、丙二醛、caspase-3以及细胞凋亡率明显升高,相关抗氧化酶的mRNA、超氧化物歧化酶及Nrf2、HO-1蛋白表达明显降低(P<0.05);与缺氧再复氧+溶剂组比较,缺氧再复氧+黄杞苷组心肌损伤标志物、活性氧、丙二醛、caspase-3以及细胞凋亡率明显下降,相关抗氧化酶的mRNA、超氧化物歧化酶及Nrf2、HO-1蛋白表达明显升高(P<0.05)。结论黄杞苷可以减轻缺氧再复氧的H9c2心肌细胞的氧化应激损伤及细胞凋亡,可能是通过Nrf2/HO-1通路发挥作用。

Objective To investigate the effect and mechanism of engeletin on hypoxia reoxygenation injury in H9c2 cells.Methods A hypoxia reoxygenation model of H9c2 cells was constructed,and H9c2 cells were randomly divided into normoxia+vehicle group,normoxia+engeletin group,hypoxia reoxygenation+vehicle group,and hypoxia reoxygenation+engeletin group.Real-time quantitative polymerase chain reaction detection of mRNA levels of related antioxidant enzymes(peroxidase 2,glutathione peroxidase 1,catalase).The kit detects levels of superoxide dismutase,myocardial injury markers,malondialdehyde,and caspase-3.Immunohistochemical staining was used to detect cellular oxidative stress levels.TdT-mediated dUTP-biotin nick end labeling assay staining was used to detect the level of cell apoptosis.Western blot was used to detect the expression levels of red blood cell nuclear factor 2 related factor 2(Nrf2)and heme oxygenase 1(HO-1)related proteins.Results There was no statistically significant difference in myocardial injury markers,oxidative stress,cell apoptosis,and Nrf2,HO-1 protein expression between the normoxia+vehicle group and the normoxia+engeletin group(P>0.05).Compared with the normoxia+vehicle group,the hypoxia reoxygenation+vehicle group showed a significant increase in myocardial injury markers,reactive oxygen species,malondialdehyde,cell apoptosis rate,and caspase-3 levels,while the mRNA levels of related antioxidant enzymes,superoxide dismutase levels,and Nrf2,HO-1 protein expression decreased significantly(P<0.05).Compared with the hypoxia reoxygenation+vehicle group,the hypoxia reoxygenation+engeletin group showed a significant decrease in myocardial injury markers,reactive oxygen species,malondialdehyde,cell apoptosis rate,and caspase-3 levels.The mRNA levels of related antioxidant enzymes,superoxide dismutase levels,and Nrf2,HO-1 protein expression were significantly increased(P<0.05).Conclusion Engeletin can alleviate oxidative stress damage and apoptosis in H9c2 cells subjected to hypoxia reoxygenation,possibly through the Nrf2/HO-1 pathway.