TAGLN2基因siRNA对脂多糖诱导的H9C2心肌细胞凋亡及氧化损伤的机制研究

Mechanism of TAGLN2 gene siRNA on lipopolysaccharide-induced apoptosis and oxidative damage of H9C2 cardiomyocytes

目的探讨抑制TAGLN2基因表达对脂多糖(LPS)诱导的H9C2心肌细胞凋亡及活性氧(ROS)水平的影响。方法将H9C2心肌细胞随机分为4组:对照组、LPS组、阴性siRNA组及TAGLN2 siRNA组,其中siRNA的转染参照LipofectamineTM2000说明。采用Western blot检测TAGLN2、核因子(NF)-κB p65、p-NF-κB p65和Bax蛋白的表达量;采用细胞计数试剂盒8(CCK8)检测细胞活力;采用流式细胞仪检测细胞凋亡率及ROS水平。结果 TAGLN2 siRNA转染后的H9C2心肌细胞TAGLN2蛋白的相对表达量降低,明显低于对照组(P<0.05)。LPS组细胞活力明显低于对照组,凋亡率、ROS水平及p-NF-κB p65和Bax蛋白的相对表达量均明显高于对照组(P<0.05),而TAGLN2 siRNA组细胞活力明显高于阴性siRNA组,凋亡率、ROS水平及p-NF-κB p65和Bax蛋白的相对表达量均明显低于阴性siRNA组(P<0.05)。结论抑制TAGLN2基因表达可降低LPS诱导的H9C2心肌细胞凋亡,可能与降低其ROS水平及下调NF-κB信号有关。

Objective To investigate the effect of that TAGLN2 gene expression was inhibited on apoptosis and reactive oxygen species( ROS) level of H9C2 cardiomyocyte induced by lipopolysaccharide (LPS). Methods H9C2 cardiomyocyte were randomly divided into 4 groups: control group, LPS group, negative siRNA group and TAGLN2 siRNA group. SiRNA was transfected referred to Lipofectamine1'1 2000. Western blotting was used to detect the expression of TAGLN2 , nuclear factor( NF)-kB p65 ,p-NF-?B p65 and Bax protein. Cell viability was detected by cell counting kit 8 ( CCK8 ). Cell apoptosis rate and ROS level were detected by flow cytometry. Results The relative expression of TAGLN2 protein in H9C2 cardiomyocyte decreased after TAGLN2 siRNA was transfect, which was significantly lower than that in control group( P <0. 05 ). Cell viability in LPS group was significantly lower than that in control group, apoptosis rate, ROS level and realtive expression of p-NF-kB p65 and Bax protein were significantly higher than those in control group(P <0. 05 ), but cell viability in TAGLN2 siRNA group was significantly higher than that in negative siRNA group, apoptosis rate, ROS level, relative expression of p-NF-kB p65 and Bax protein were significantly lower than those in negative siRNA group( P <0. 05). Conclusion Inhibition of TAGLN2 gene expression can reduce the apoptosis of H9C2 cardioinyocyte induced by LPS, which may be associated with decreased ROS level and down-regulation of NF-kB signal.