miRNA-130a-3p通过调控NF-κB信号通路影响损伤心肌细胞

Effects of miRNA-130a-3p on cardiomyocyte injury through regulating NF-κB signaling pathway

目的探讨miRNA-130a-3p在LPS诱导的心肌细胞损伤中的作用及可能机制。方法H9C2心肌细胞分为4组,即正常对照组、LPS模型组、miRNA阴性对照组、miRNA-130a-3p mimics组,利用RT-qPCR检测miRNA-130a-3p mRNA表达水平,CCK-8检测细胞活性,ELISA检测培养液中TNF-α、IL-6、IL-1β含量,Western blot检测NF-κBp65、IκBα、Bax、Bcl-2、Cleaved-Caspase-3蛋白表达水平。结果RT-qPCR结果显示LPS模型细胞中miRNA-130a-3p mRNA水平显著低于正常对照组;与正常对照组相比较,LPS组细胞活性显著降低,而与LPS组相比较,miRNA-130a-3p mimics组细胞活性显著升高;与正常对照组相比较,LPS组细胞培养液中TNF-α、IL-6、IL-1β含量显著升高,而与LPS组相比较,miRNA-130a-3p mimics组细胞培养液中TNF-α、IL-6、IL-1β含量显著降低;与正常对照组相比较,LPS组细胞中NF-κBp65、Bax、Cleaved-Caspase-3蛋白表达量显著升高,IκBα与Bcl-2蛋白表达量显著降低,而与LPS组相比较,miRNA-130a-3p mimics组细胞中NF-κBp65、Bax及Cleaved-Caspase-3蛋白表达量显著降低,IκBα与Bcl-2蛋白的表达量显著升高。结论miRNA-130a-3p可通过抑制NF-κB信号通路激活,抑制细胞凋亡,提高细胞活性,减少炎性因子释放,从而减轻LPS诱导的心肌细胞损伤。

Objective To investigate the role and possible mechanism of miRNA-130a-3p in cardiomyocyte injury induced by LPS.Methods H9C2 cells were divided into the following four groups:a normal control group,a LPS model group,a miRNA negative control group and a miRNA-130a-3p mimics group.The expression level of miRNA-130a-3p mRNA in each group was detected by RT-qPCR.The CCK-8 assay was used to detect the cell activity in each group.The contents of TNF-α,IL-6 and IL-1βwere detected by ELISA assay.Western blot was used to detect the protein expression levels of NF-κBp65,IκBα,Bax,Bcl-2 and Cleaved-Caspase-3.Results The results of RT-qPCR showed that the level of miRNA-130a-3p mRNA in the LPS model group was significantly lower than that in normal control group.The results of CCK-8 showed that the cell activity of LPS group was significantly lower than that of normal control group,while compared with the LPS group,the cell activity in miRNA-130a-3p mimics group increased significantly.The results of ELISA showed that the contents of TNF-α,IL-6 and IL-1βin cell culture medium of LPS group were significantly higher than those of normal control group,while compared with the LPS group,the contents of TNF-α,IL-6 and IL-1βin cell culture medium of miRNA-130a-3p mimics group decreased significantly.The results of Western blot showed that compared with normal control group,the protein expression levels of NF-κBp65,Bax and Cleaved-Caspase-3 increased significantly in the LPS group,while the protein expression levels of IκBαand Bcl-2 decreased significantly.Compared with the LPS group,the protein expression levels of NF-κBp65,Bax and Cleaved-Caspase-3 decreased significantly,while the protein expression levels of IκBαand Bcl-2 increased significantly in miRNA-130a-3p mimics group.Conclusions miRNA-130A-3P can inhibit cell apoptosis,improve cell activity and reduce the release of inflammatory factors,so as to alleviate cardiomyocyte injury induced LPS,through inhibiting the activation of NF-κB signaling pathway.