黄芪多糖联合lncRNA TUG1沉默对缺血再灌注人心脏微血管内皮细胞凋亡的影响

Effects of Astragalus Polysaccharide Combined with lncRNA TUG1 Silencing on the Apoptosis of Human Cardiac Microvascular Endothelial Cells after Ischemia-reperfusion

目的:探讨黄芪多糖联合长链非编码RNA(lncRNA)牛磺酸调节基因1(TUG1)沉默对缺血再灌注人心脏微血管内皮细胞凋亡的影响。方法:将心脏微血管内皮细胞分成对照组、模型组、sh-NC组、sh-TUG1组、黄芪多糖组、黄芪多糖+sh-TUG1组,流式细胞术检测凋亡,Western Blotting方法检测音速豪猪蛋白(SHH)表达,ELISA法检测肿瘤坏死因子-α(TNF-α)含量,DCFH-DA法检测活性氧(ROS)含量。结果:与对照组比较,模型组心脏微血管内皮细胞凋亡率升高,TNF-α增多,ROS升高,SHH蛋白表达降低(均P<0.05);与sh-NC组比较,sh-TUG1组心脏微血管内皮细胞凋亡率降低,TNF-α减少,ROS含量降低,SHH蛋白表达升高(均P<0.05);与模型组比较,黄芪多糖组心脏微血管内皮细胞凋亡率降低,TNF-α减少,ROS降低,SHH蛋白表达升高(均P<0.05);与sh-TUG1组、黄芪多糖组比较,黄芪多糖+sh-TUG1组心脏微血管内皮细胞凋亡率降低,TNF-α减少,ROS含量降低,SHH蛋白表达升高(均P<0.05)。结论:黄芪多糖联合TUG1基因沉默能减少缺血再灌注人心脏微血管内皮细胞凋亡,减轻氧化损伤,抑制炎症介质分泌,其机制可能与激活SHH信号通路有关。

Objective:To explore the effects of astragalus polysaccharide combined with long non-coding RNA(lncRNA)and taurine up-regulated gene 1(TUG1)on the apoptosis of human cardiac microvascular endothelial cells after ischemia-reperfusion.Methods:Cardiac microvascular endothelial cells were divided into a control group,a model group,sh-NC group,sh-TUG1 group,astragalus polysaccharide group,astragalus polysaccharide+sh-TUG1 group.Flow cytometry was used to detect apoptosis,and Western Blotting method was used to detect sonic hedgehog(SHH)expression.ELISA method to detect tumor necrosis factor-α(TNF-α)content,and DCFH-DA method was used to detect reactive oxygen species(ROS)content.Results:Compared with the control group,cardiac microvascular endothelial cells in the model group apoptosis rate increased,TNF-αincreased,ROS content increased,SHH protein expression decreased(P s<0.05);compared with sh-NC group,cardiac microvascular endothelial cells of sh-TUG1 group apoptosis rate decreased,TNF-αdecreased,ROS decreased,SHH protein expression increased(all P s<0.05);compared with the model group,the apoptosis rate of cardiac microvascular endothelial cells in the astragalus polysaccharide group decreased,TNF-αdecreased,ROS decreased,SHH protein expression increased(all P s<0.05);compared with the sh-TUG1 and the astragalus polysaccharide group,the apoptosis rate of cardiac microvascular endothelial cells in the astragalus polysaccharide+sh-TUG1 group decreased,TNF-αdecreased,ROS decreased,SHH protein expression increased(P s<0.05).Conclusion:Astragalus polysaccharide combined with TUG1 gene silencing can reduce the apoptosis of ischemia-reperfusion human cardiac microvascular endothelial cells,reduce oxidative damage,and inhibit the secretion of inflammatory factors,the mechanism may be related to the activation of SHH signaling pathway.