体外转染Klotho基因对H9c2(2-1)大鼠心肌细胞缺血再灌注的影响

摘要

目的 研究转染Klotho基因对H9c2(2-1)大鼠心肌细胞缺血再灌注的影响.方法 培养H9c2(2-1)大鼠心肌细胞,建立模拟缺血再灌注模型,Fermentas转染试剂介导小鼠Klotho基因转染H9c2(2-1)大鼠心肌细胞.实验分4组:对照组(Control组)、缺血再灌注组(I-R组)、转染Klotho基因组(Klotho组)、转染Fermentas转染试剂空载体组(Vehicle control组).各组进行缺血再灌注后RT-PCR及免疫荧光法检测Klotho mRNA 及Klotho蛋白表达,MTT法检测细胞活性,测定上清液乳酸脱氢酶(LDH)、丙二醛(MDA)及Klotho含量.结果 Klotho组较I-R组和Vehicle control 组Klotho mRNA、Klotho蛋白表达明显增加,LDH、MDA明显降低,细胞活性高(p＜0.01).结论 Fermentas转染试剂介导的Klotho基因转染H9c2 (2-1)大鼠心肌细胞可保护H9c2(2-1)大鼠心肌细胞、减轻H9c2(2-1)大鼠心肌细胞缺血再灌注损伤.%Objective To explore the effects and mechanisms of Fermentas transfection reagent mediated transfection of Klotho gene on ischemia-reperfusion (I/R) injury in H9c2(2-1) rat cardiomyocytes in vitro. Methods H9c2(2-1) rat cardiomyocytes were cultivated and the model of I/R was established Fermentas transfection reagent mediated Klotho gene was transfected into H9c2 (2-1) rat cardiomyocytes. H9c2(2-1) rat cardiomyocytes were randomly divided into control, I/R,Klotho and vehicle control groups. After the model was established , the expressions of Klotho mRNA and protein were tested by RT-PCR, immunofluorescence and ELISA. MIT was used to investigate the cell activity. Levels of lactate dehydrogenase ( LDH), malondialdehyde ( MDA) in the medium were tested. Results In comparison with I/R and vehicle control groups, Klotho mRNA and protein expression and cell activity in Klotho transfected group were significantly in- ■ creased ( P <0.01) , however, the levels of LDH and MDA in Klotho transfected group were decreased obviously (P <0. 01). Conclusions Fermentas transfection reagent mediated transfection of Klotho gene can protect cultured H9c2(2-1) rat cardiomyocytes against I/R injury.