长链非编码RNA NRON促进NFATc3磷酸化减轻心房纤维化的机制研究

LncRNA NRON alleviates atrial fibrosis via promoting NFATc3 phosphorylation

目的探讨长链非编码RNA(LncRNA)NRON在心房纤维化过程中的作用。方法使用qRT-PCR检测NRON在心房组织中的表达水平。通过蛋白质印迹法测定胶原蛋白Ⅰ、胶原蛋白Ⅲ、NFATc3和pNFATc3的蛋白水平。再进行免疫组织化学测定以观察心房组织中胶原Ⅰ的表达和分布。通过波形蛋白/肌钙蛋白免疫荧光染色对心房成纤维细胞进行鉴定。继而用CCK-8分析检测成纤维细胞增殖。通过HE染色观察心脏组织的形态变化,最后Masson染色检测心肌纤维化。结果NRON可以抑制成纤维细胞增殖与胶原Ⅰ和胶原Ⅲ的表达,激活NFATc3和核素导入。过表达NRON可以促进NFATc3的磷酸化,从而抑制小鼠心脏纤维化进程。结论LncRNA NRON促进NFATc3磷酸化,抑制活化T细胞因子(NFAT)的活性,从而抑制心房组织的纤维化进程。

Objective To explore the specific mechanism of LncRNA NRON affecting NFAT activity and progression of atrial fibrosis.Methods The expression level of NRON in atrial tissue was detected by qRT-PCR.Western blot was used to determine the protein levels of collagenⅠ,collagenⅢ,NFATc3 and pNFATc3.Immunohistochemistry was performed to observe the expression and distribution of collagenⅠin atrial tissue.Atrial fibroblasts were identified by vimentin/troponin immunofluorescence staining.Then the proliferation of fibroblasts was detected by CCK-8 analysis.HE staining was used to observe the morphological changes of cardiac tissue,and finally the myocardial fibrosis was detected by Masson staining.Results NRON inhibited fibroblast proliferation,and the expression of collagenⅠand collagenⅢ,activated NFATc3 and nuclide entry.Overexpression of NRON inhibited the cardiac fibrosis induced by NFAT in mice.Conclution The data from this study suggest that NRON alleviates atrial fibrosis by promoting NFATc3 phosphorylation.