The long noncoding RNA NR_045363 involves cardiomyocyte apoptosis and cardiac repair via p53 signal pathway
The long noncoding RNA NR_045363 involves cardiomyocyte apoptosis and cardiac repair via p53 signal pathway
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长非编码RNA NR_045363通过p53信号通路参与心肌细胞凋亡和心脏修复
DOI：
10.1002/cbin.11374
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发表时间：
2020-06-10
影响因子：
3.9
通讯作者：
Chu, Maoping
中科院分区：
文献类型：
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作者：
Chen, Xianda;Wang, Jue;Chu, Maoping
长非编码RNA(LncRNAs)参与多种生物学行为，包括调节细胞分化、增殖和凋亡。研究人员此前证实，高度保守的IncRNA NR_045363控制心肌细胞(CM)的增殖和心脏修复。本研究旨在探讨NR_045363对心肌细胞凋亡的影响。7日龄小鼠永久结扎冠状动脉前降支(LAD)，用实时定量聚合酶链式反应(QRT-PCR法)检测NR045363的表达。心肌梗死组NR_045363的表达在术后1周明显高于假手术组。以NR_045363为靶向的LncRNA Smart消音剂下调原代培养的CMS中NR_045363的表达，并用末端脱氧核苷酸酶介导的缺口末端标记和Annexin-V/PI双重染色分析CMS的凋亡。上述结果表明，NR_045363基因敲除可显著促进CMS的凋亡。为了探讨其内在机制，进行了RNA测序(RNA-SEQ)，并用独创性通径分析(IPA)对RNA-SEQ结果进行了分析。序列分析表明，在NR_045363基因敲除的细胞质雄性不育系中，共有2,291个基因表达上调或下调，IPA分析表明肿瘤蛋白53(P53)是上游调控基因。在体内，通过AAV9系统过表达的NR_045363改善了7日龄小鼠心肌梗死后的心功能，并抑制了心肌细胞的凋亡。这些结果提示，NR_045363参与了心肌细胞的凋亡，而NR_045363的过表达通过抑制P53途径减轻了心肌细胞的凋亡，从而对心脏修复起到积极作用。
Long noncoding RNAs (lncRNAs) can participate in various biological behaviors, including regulating cell differentiation, proliferation, and apoptosis. The investigators have previously confirmed that highly conserved lncRNA NR_045363 controls cardiomyocyte (CM) proliferation and cardiac repair. The present study investigates the effects of NR_045363 on CM apoptosis. Seven-day-old mice were subjected to permanent left anterior descending coronary artery ligation (LAD), and the NR_045363 expression was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of NR_045363 in the MI group significantly exceeded the Sham group during the first week after the operation. The NR_045363 expression was knocked down in primary cultured CMs using an NR_045363-targeting lncRNA Smart silencer, and the apoptosis of CMs was analyzed by terminal-deoxynucleoitidyl transferase mediated nick end labeling and Annexin-V/PI double staining. These present results indicate that the NR_045363 knockdown significantly promoted the apoptosis of CMs. In order to investigate the underlying mechanism, RNA-sequencing (RNA-seq) was performed, and ingenuity pathway analysis (IPA) was used to analyze the RNA-seq results. The RNA-seq data revealed that a total of 2,291 genes were upregulated or downregulated in NR_045363 knockdown CMs, and the IPA analysis indicated that tumor protein 53 (p53) was the upstream regulator. In vivo, the NR_045363 overexpression through the AAV9 system improved the heart function after MI in 7-day-old mice and inhibited the CM apoptosis. These data suggest that NR_045363 is involved in CM apoptosis and that NR_045363 overexpression exerts positive effects on cardiac repair by alleviating CM apoptosis through the inhibition of the p53 pathway.