LncRNA 1700020I14Rik/miR-297a/CGRP axis suppresses myocardial cell apoptosis in myocardial ischemia-reperfusion injury
LncRNA 1700020I14Rik/miR-297a/CGRP axis suppresses myocardial cell apoptosis in myocardial ischemia-reperfusion injury
复制标题
LncRNA 1700020I14Rik/miR-297a/CGRP轴抑制心肌缺血再灌注损伤中的心肌细胞凋亡
DOI：
10.1016/j.molimm.2020.03.015
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发表时间：
2020-06-01
影响因子：
3.6
通讯作者：
Jiang, Zhengming
中科院分区：
文献类型：
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作者：
Hu, Fudong;Yang, Jinhua;Jiang, Zhengming
背景：长链非编码RNA（lncRNA）与多种人类疾病密切相关，但其在心肌损伤中的作用尚未完全阐明。本研究发现lncRNA 1700020I14Rik在心肌损伤组织中表达显着下调，并探讨lncRNA 1700020I14Rik调节心肌细胞损伤的机制。方法：建立心肌缺血再灌注（I/R）损伤和心肌细胞缺氧/复氧（H/R）模型。 建立损伤并通过 qRT-PCR 或蛋白质印迹分析 1700020I14Rik、miR-297a 或 CGRP 的表达。此外，通过TUNEL染色评估心肌细胞凋亡，并通过LDH细胞毒性检测试剂盒测量小鼠血浆样品或心肌细胞培养上清液中LDH的浓度。此外，通过MTT法评估H/R处理后心肌细胞存活率的差异，并在用1700020I14Rik或miR-297a过表达或沉默的HL-1细胞中观察CGRP表达。此外，通过双荧光素酶报告基因分析分析miR-297a对1700020I14Rik和CGRP表达的调节功能。结果：在心肌I/R损伤和心肌细胞H/R损伤模型中，1700020I14Rik和CGRP表达异常下调，而miR-297a表达上调。 TUNEL染色显示，心肌I/R损伤模型中心肌细胞凋亡率明显升高。此外，心肌细胞损伤后，小鼠血浆样品或心肌细胞培养上清液中LDH的浓度显着升高。 MTT法检测发现H/R处理心肌细胞后细胞存活率下降。此外，过表达1700020I14Rik或敲低miR297a可上调CGRP蛋白水平，而干扰1700020I14Rik或过表达miR297a则产生相反的结果。进一步的研究证实lncRNA 1700020I14Rik/miR-297a/CGRP轴抑制心肌I/R损伤中的心肌细胞凋亡。结论：我们的结果表明1700020I14Rik在心肌损伤组织中异常下调。深入研究表明1700020I14Rik/miR-297a/CGRP轴在心肌I/R损伤中抑制心肌细胞凋亡。
Background: Long non-coding RNAs (lncRNAs) are closely related to various human diseases, but their role in myocardial injury has not been fully elucidated. In the current study, we found that the expression of lncRNA 1700020I14Rik was significantly down-regulated in myocardial injury tissues and the underlying mechanism by which lncRNA 1700020I14Rik regulated myocardial cell injury was investigated.Methods: The model of myocardial ischemia-reperfusion (I/R) injury and myocardial cells hypoxia/reoxygenation (H/R) injury were established and the expression of 1700020I14Rik, miR-297a or CGRP was analyzed by qRT-PCR or Western blot. Moreover, myocardial cell apoptosis was assessed by TUNEL staining and the concentration of LDH in the mouse plasma sample or myocardial cell culture supernatant was measured by the LDH cytotoxicity test kit. Furthermore, the differences of myocardial cell survival rate after H/R treatment were assessed by MTT assay and the observation of CGRP expression was performed in HL-1 cells overexpressed or silenced with 1700020I14Rik or miR-297a. In addition, the regulating function of miR-297a on 1700020I14Rik and CGRP expression was analyzed by a dual luciferase reporter assay.Results: The expressions of 1700020I14Rik and CGRP were abnormally down-regulated in a model of myocardial I/R injury and myocardial cells H/R injury, while miR-297a was up-regulated. By TUNEL staining, the apoptotic rate of myocardial cells in the model of myocardial I/R injury was significantly increased. Furthermore, the concentrations of LDH in the mouse plasma sample or myocardial cell culture supernatant were significantly increased after myocardial cell injury. By MTT assay, the survival rate of cells was decreased after myocardial cells were treated with H/R. In addition, overexpression of 1700020I14Rik or knockdown of miR297a could up-regulate CGRP protein level, while interference with 1700020I14Rik or overexpression of miR297a produced the opposite result. Further study confirmed that lncRNA 1700020I14Rik/miR-297a/CGRP axis suppressed myocardial cell apoptosis in myocardial I/R injury.Conclusion: Our results indicated that 1700020I14Rik was abnormally down-regulated in myocardial injury tissues. In-depth studies manifested that 1700020I14Rik/miR-297a/CGRP axis suppressed myocardial cell apoptosis in myocardial I/R injury.