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。
Abstract
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-1β were 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-1β in 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-1β in 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 IκBα and 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 IκBα and 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.
关键词
微小RNA-130a-3p； /
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NF-κB信号通路； /
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细胞损伤； /
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细胞凋亡
Key words
miRNA-130a-3p; /
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NF-κB signaling pathway; /
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Cell injury; /
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Apoptosis
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基金
河南省科技厅科技攻关项目（212102310681）；2021年度河南省高等学校青年骨干教师培养计划（2021GGJS200）
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