微小 RNA1283 通过靶向 GADD45A 并抑制 JNK 和 p38MAPK 信号通路缓解低氧/复氧诱导的心肌细胞损伤。
MicroRNA 1283 alleviates cardiomyocyte damage caused by hypoxia /reoxygenation via targeting GADD45A and inactivating the JNK and p38 MAPK signaling pathways.
机构信息
Department of Biochemistry and Molecular Biology, School of Basic Medicine, Jiamusi University, Jiamusi, Heilongjiang, China
Department of Biochemistry and Molecular Biology, Jiamusi University, Jiamusi, Heilongjiang, China
出版信息
Kardiol Pol. 2021 Feb 25;79(2):147-155. doi: 10.33963/KP.15696. Epub 2020 Dec 3.
BACKGROUND
Clarifying themolecular mechanism and identifying markers of myocardial ischemia/reperfusion injury is crucial for the treatment of acute myocardial infarction.
AIMS
This study aimed to investigate the roles and underlying regulatory mechanisms of microRNA 1283 (miR-1283) and GADD45A in cardiomyocytes injured by hypoxia /reoxygenation (H/R).
METHODS
Bioinformatic analyses were used to determine the expression of GADD45A and miR-1283 based on various datasets from the Gene Expression Omnibus database. Human embryonic cardiomyocytes were subjected to H/R to construct in vitro models. Real -time quantitative polymerase chain reaction and Western blot were used to detect mRNA and protein expression levels, respectively. The binding sites between miR-1283 and GADD45A were predicted by the TargetScan software and verified using dual luciferase reporter assays. Cell viability and apoptosis were detected with the use of Cell Counting Kit 8 and flow cytometry assays.
RESULTS
GADD45A and miR-1283 were upregulated or downregulated in myocardial infarction, respectively. MicroRNA 1283 expression was decreased in cardiomyocytes after H/R treatment. H/R treatment reduced cardiomyocyte viability and enhanced apoptosis, and these effects were abated by transfection of a miR1283 mimic and strengthened by transfection of a miR-1283 inhibitor. MicroRNA 1283 bound to the 3' untranslated region of GADD45A and decreased the levels of GADD45A, which inhibited proliferation and promoted apoptosis in H/R -induced cardiomyocyte injury. Reintroduction of GADD45A attenuated the effect of miR-1283 on the viability and apoptosis of cardiomyocytes in H/R models. The JNK and p38 MAPK signaling pathways were regulated by the miR-283-GADD45A axis.
CONCLUSIONS
The miR-1283-GADD45A axis may protect against H/R -induced cardiomyocyte injury by suppressing the JNK and p38 MAPK pathways.
背景
阐明心肌缺血/再灌注损伤的分子机制并鉴定其标志物，对于急性心肌梗死的治疗至关重要。
目的
本研究旨在探讨微小 RNA 1283（miR-1283）和生长停滞及 DNA 损伤诱导蛋白 45A（GADD45A）在缺氧/复氧（H/R）诱导的心肌细胞损伤中的作用及其潜在调控机制。
方法
基于基因表达综合数据库中的各种数据集，采用生物信息学分析方法确定 GADD45A 和 miR-1283 的表达。将人胚胎心肌细胞进行 H/R 处理，构建体外模型。实时定量聚合酶链反应和 Western blot 分别用于检测 mRNA 和蛋白表达水平。TargetScan 软件预测 miR-1283 与 GADD45A 之间的结合位点，并通过双荧光素酶报告基因实验进行验证。采用细胞计数试剂盒 8 和流式细胞术检测细胞活力和凋亡。
结果
GADD45A 和 miR-1283 在心肌梗死中分别上调或下调。H/R 处理后心肌细胞中 miR-1283 的表达降低。H/R 处理降低了心肌细胞活力并增强了细胞凋亡，而过表达 miR-1283 模拟物可减轻这些作用，而过表达 miR-1283 抑制剂则可增强这些作用。miR-1283 与 GADD45A 的 3'非翻译区结合，降低了 GADD45A 的水平，从而抑制了 H/R 诱导的心肌细胞损伤中的增殖并促进了细胞凋亡。GADD45A 的再导入减弱了 miR-1283 对 H/R 模型中心肌细胞活力和凋亡的影响。miR-1283-GADD45A 轴通过调节 JNK 和 p38 MAPK 信号通路来发挥作用。
结论
miR-1283-GADD45A 轴可能通过抑制 JNK 和 p38 MAPK 通路来保护心肌细胞免受 H/R 诱导的损伤。
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