Abstract

Long noncoding RNAs (lncRNAs) have been considered as crucial regulators of acute myocardial infarction (AMI). In this study, to analyze the effect of differentiation antagonizing nonprotein coding RNA (DANCR) of lncRNA on cardiomyocyte damage in AMI, cardiomyocyte injury was induced by oxygen-glucose deprivation (OGD). Cell counting kit-8 (CCK-8) assay and flow cytometry were used to assess cell viability and apoptosis, respectively. Quantitative real-time PCR was used to measure the expression levels of DANCR and miR-19a-3p. Bioinformatics analysis and luciferase gene reporter assay were utilized to explore the relationship among DANCR, miR-19a-3p, and mitogen-activated protein kinase 1 (MAPK1). CCK-8 and TUNEL assays were used to explore the effects of DANCR alone or plus miR-19a-3p on the viability and apoptosis of OGD/R-exposed HL-1 cells. Western blot analysis was used to detect changes in the MAPK1/ERK1/2 pathway in HL-1 cells. We found that DANCR expression and miR-19a-3p level are negatively correlated as DANCR expression is increased, while miR-19a-3p level is decreased in AMI patients’ serum and OGD/R-exposed HL-1 cells. DANCR knockdown increased miR-19a-3p level, and miR-19a-3p inhibition increased DANCR expression. Moreover, DANCR directly binds to miR-19a-3p. DANCR knockdown reduced viability but induced apoptosis in OGD/R-exposed HL-1 cells, while miR-19a-3p inhibition weakens these effects. Furthermore, MAPK1 is a target of miR-19a-3p. miR-19a-3p overexpression decreases MAPK1 and ERK1/2 in HL-1 cells, while miR-19a-3p inhibition increases MAPK1 and ERK1/2 in HL-1 cells. Moreover, DANCR knockdown reduces myocardium apoptosis in mice with the left anterior descending artery ligated. DANCR knockdown effectively restores myocardial cell apoptosis by regulating the miR-19a-3p/MAPK1/ERK1/2 axis.

中文翻译：

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抑制 lncRNA DANCR 通过 miR-19a-3p/MAPK1 轴减轻氧-葡萄糖剥夺诱导的心肌细胞损伤

摘要

长链非编码 RNA (lncRNA) 被认为是急性心肌梗死 (AMI) 的关键调节因子。本研究为分析拮抗lncRNA分化的非蛋白编码RNA（DANCR）对AMI心肌细胞损伤的影响，采用氧糖剥夺（OGD）诱导心肌细胞损伤。细胞计数试剂盒 8 (CCK-8) 测定和流式细胞仪分别用于评估细胞活力和细胞凋亡。定量实时 PCR 用于测量 DANCR 和 miR-19a-3p 的表达水平。利用生物信息学分析和荧光素酶基因报告基因分析来探索 DANCR、miR-19a-3p 和丝裂原活化蛋白激酶 1 (MAPK1) 之间的关系。CCK-8 和 TUNEL 测定用于探索 DANCR 单独或加 miR-19a-3p 对 OGD/R 暴露的 HL-1 细胞的活力和凋亡的影响。Western印迹分析用于检测HL-1细胞中MAPK1/ERK1/2通路的变化。我们发现，随着 DANCR 表达增加，DANCR 表达和 miR-19a-3p 水平呈负相关，而 AMI 患者血清和 OGD/R 暴露的 HL-1 细胞中 miR-19a-3p 水平降低。DANCR 敲低增加了 miR-19a-3p 水平，而 miR-19a-3p 抑制增加了 DANCR 表达。此外，DANCR 直接与 miR-19a-3p 结合。DANCR 敲低降低了 OGD/R 暴露的 HL-1 细胞的活力，但诱导了细胞凋亡，而 miR-19a-3p 抑制减弱了这些作用。此外，MAPK1 是 miR-19a-3p 的靶标。miR-19a-3p 过表达会降低 HL-1 细胞中的 MAPK1 和 ERK1/2，而 miR-19a-3p 抑制会增加 HL-1 细胞中的 MAPK1 和 ERK1/2。而且，DANCR 敲低可减少左前降支结扎小鼠的心肌细胞凋亡。DANCR 敲低通过调节 miR-19a-3p/MAPK1/ERK1/2 轴有效地恢复心肌细胞凋亡。