lncRNA SLC8A1-AS1在脓毒症大鼠心肌组织中的表达及其对心肌细胞凋亡和炎症因子分泌的影响

Expression of lncRNA SLC8A1-AS1 in myocardial tissue of sepsis rats and its effect on myocardial cell apoptosis and secretion of inflammatory factors

目的研究lncRNA SLC8A1-AS1在脓毒症大鼠心肌组织中表达及其对心肌细胞凋亡和炎症因子分泌的影响。方法qRT-聚合酶链反应(PCR)检测脓毒症大鼠心肌组织中SLC8A1-AS1表达水平。心肌H9C2细胞分成Control组(正常培养)、脂多糖(LPS)组(LPS诱导处理)、Vector+LPS组(转染阴性对照载体,LPS诱导处理)、SLC8A1-AS1+LPS组(转染SLC8A1-AS1过表达载体,LPS诱导处理)、SLC8A1-AS1+LPS+佛波酯(PMA)组[转染SLC8A1-AS1过表达载体,核转录因子(NF)-κB信号激活剂和LPS诱导处理]。四甲基偶氮唑蓝(MTT)比色法检测细胞增殖变化,流式细胞术检测细胞凋亡水平,酶联免疫吸附试验(ELISA)检测细胞分泌的白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α水平,Western印迹检测酶切含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-3、p65蛋白表达水平变化。结果脓毒症大鼠心肌组织中SLC8A1-AS1表达水平显著降低(P<0.05)。与Control组比较,LPS组心肌细胞增殖活性显著降低,细胞凋亡率显著升高,细胞分泌的IL-1β、IL-6、TNF-α显著增多,酶切Caspase-3、p65蛋白表达显著增多(均P<0.05)。与Vector+LPS组比较,SLC8A1-AS1+LPS组心肌细胞增殖活性显著升高,细胞凋亡率显著降低,细胞分泌的IL-1β、IL-6、TNF-α显著减少,酶切Caspase-3、p65蛋白表达显著减少(均P<0.05)。与SLC8A1-AS1+LPS组比较,SLC8A1-AS1+LPS+PMA组心肌细胞增殖活性显著降低,细胞凋亡率显著升高,细胞分泌的IL-1β、IL-6、TNF-α显著增多,酶切Caspase-3、p65蛋白表达显著增多(均P<0.05)。结论SLC8A1-AS1在脓毒症大鼠心肌组织中表达下降,上调SLC8A1-AS1通过抑制NF-κB信号减少心肌细胞凋亡和分泌炎症因子。

Objective To investigate the expression of lncRNA SLC8 A1-AS1 in myocardial tissue of sepsis rats and its effect on myocardial cell apoptosis and secretion of inflammatory factors.Methods qRT-PCR method was used to detect the expression level of SLC8 A1-AS1 in myocardial tissue of sepsis rats.Cardiac H9 C2 cells were divided into Control group,LPS group(LPS was used to induce treatment),Vector+LPS group(negative control vector was transfected,LPS was used to induce treatment),SLC8 A1-AS1+LPS group(SLC8 A1-AS1 overexpression vector was transfected,LPS was used to induce treatment),SLC8 A1-AS1+LPS+PMA group(transfected with SLC8 A1-AS1 overexpression vector,NF-κB signal activator and LPS induction treatment).Cell proliferation was detected by MTT colorimetry,cell apoptosis was detected by flow cytometry,the levels of IL-1β,IL-6 and TNF-α secreted by cells were detected by ELISA,the expression of C-Caspase-3 and p65 protein in cells level changes were detected by Western blot.Results The expression level of SLC8 A1-AS1 in myocardium of sepsis rats was significantly decreased(P<0.05).Compared with the Control group,the proliferation activity of cardiomyocytes in the LPS group was significantly decreased,the apoptosis rate was significantly increased,the secretion of IL-1β,IL-6 and TNF-α were significantly increased,and the expression of C-Caspase-3 and p65 proteins in the cells were significantly increased(P<0.05).Compared with the Vector+LPS group,the proliferation activity of cardiomyocytes in the SLC8 A1-AS1+LPS group was significantly increased,the apoptosis rate was significantly decreased,the secretion of IL-1β,IL-6 and TNF-α were significantly decreased,and the expression of C-Caspase-3 and p65 proteins in the cells were significantly decreased(P<0.05).Compared with the SLC8 A1-AS1+LPS group,the proliferation activity of cardiomyocytes in the SLC8 A1-AS1+LPS+PMA group was significantly decreased,the apoptosis rate was significantly increased,the secretion of IL-1β,IL-6 and TNF-α were significantly increased,and the expression of C-Caspase-3 and p65 proteins in the cells were significantly increased(P<0.05).Conclusions The expression of SLC8 A1-AS1 is decreased in myocardial tissue of sepsis rats,up-regulation of SLC8 A1-AS1 could reduce cardiomyocyte apoptosis and secrete inflammatory factors by inhibiting NF-κB signaling.