LncRNA3215-ceRNA-miR1594调控肌钙蛋白T参与鸡缺硒性心肌细胞凋亡的作用研究

Myocardium is one of the target organs of selenium deficiency. In our previous study, we found that LncRNA-3215 upregulated in chicken heart induced by Se deficiency by whole transcriptome resequencing.The possible related miR-1594 is significantly down-regulated. We also predict that their target gene is troponin T (cTnT). However, it is not clear whether LncRNA can act as ceRNA to participate in the regulation on cTnT by miRNA and its mechanism in Se-deficient myocardial injury in chickens still unclear.. Base on the cases model of myocardial damage induced by Se deficiency in chickens. To detect of LncRNA-3215, miR-1594, intracellular calcium homeostasis, apoptosis-related genes and proteins expression in myocardial tissue, combined with previous bioinformatics results to identify the target gene cTnT and the possible signal transduction pathways in Se-induced cardiomyocyte damage in chickens. On this basis, chicken cardiomyocytes were cultured in vitro to established LncRNA-3215 and miR-1594 inhibition and overexpression models and LncRNA-3215 and cTnT overexpressed chicken embryo models, respectively. To detect the intracellular calcium homeostasis, apoptosis-related genes and proteins expression in cardiomyocytes and chicken embryo models to illuminate the mechanism of LncRNA and miRNA regulatory networks in apoptosis in myocardial damage induced by selenium deficiency.

心肌是硒缺乏的靶器官之一。课题组前期应用组学技术发现，LncRNA-3215在鸡缺硒性心肌损伤过程中显著上调，其可能作用miR-1594显著下调，且预测共同靶基因为肌钙蛋白T（cTnT），但lncRNA是否可作为ceRNA参与miRNA调控cTnT及其在鸡缺硒性心肌损伤中的作用机制尚不清楚。.本课题在建立鸡缺硒性心肌损伤模型基础上，通过对心肌组织LncRNA-3215、miR-1594及细胞内钙稳态和细胞凋亡相关通路的检测，结合前期生物信息学结果，对靶基因cTnT进行鉴定并确定其在鸡缺硒性心肌细胞损伤中的可能信号转导通路，在此基础上通过体外培养鸡心肌细胞，分别建立LncRNA-3215、miR-1594抑制及过表达模型以及LncRNA-3215、cTnT基因过表达鸡胚模型，检测细胞内钙稳态及细胞凋亡相关通路，阐明LncRNA和miRNA调控网络在鸡缺硒性心肌损伤性凋亡中的作用机制。

心肌是硒缺乏的靶器官之一。课题组前期应用组学技术发现，LncRNA-3215在鸡缺硒性心肌损伤过程中显著上调，其可能作用miR-1594显著下调，且预测共同靶基因为肌钙蛋白T（cTnT），然而在缺硒引发的心血管疾病中是否存在ceRNA调控机制目前仍知之甚少。.本课题在建立鸡缺硒性心肌和心肌细胞损伤模型的基础上，通过对心肌组织LncRNA3215、miR-1594及细胞凋亡、细胞内钙稳态、扩张性心肌病相关通路的检测，结合前期生物信息学结果，对miR-1594的靶基因TNNT2进行鉴定并确定其在鸡缺硒性心肌细胞损伤中可能的信号转导通路，在此基础上通过体外培养鸡心肌细胞以及鸡胚体内注射，分别建立LncRNA3215、miR-1594、TNNT2敲低及过表达模型以及LncRNA3215、TNNT2基因敲低鸡胚模型，检测细胞凋亡、细胞内钙稳态及扩张性心肌病相关通路基因及蛋白表达，结果表明，LncRNA3215可以通过靶向miR-1594调节TNNT2的表达，进而形成ceRNA调控机制，该机制在缺硒性心肌损伤过程中发挥重要作用。硒缺乏通过LncRNA3215-miR-1594-TNNT2轴激活氧化应激，进而诱发细胞凋亡。LncRNA3215的抑制可以通过LncRNA3215-miR-1594-TNN2途径介导硒缺乏引起的鸡心肌细胞钙超载。LncRNA3215通过在竞争性抑制miR-1594调节TNNT2的表达，进而保护硒缺乏引起的扩张型心肌病。阐明LncRNA和miRNA调控网络在鸡缺硒性心肌损伤中的作用机制。

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