Department of Cardiovascular Diseases, First Affiliated Hospital, Heilongjiang University of Chinese Medicine, Harbin, PR China.
Department of Cardiology, General Hospital of Heilongjiang Agricultural Reclamation Bureau, Harbin, PR China.
Toxicol Appl Pharmacol. 2021 Mar 1;414:115408. doi: 10.1016/j.taap.2021.115408. Epub 2021 Jan 19.
This study proposed to investigate the function of miR-19a/ACSL axis in hypoxia/reoxygenation (H/R)-induced myocardial injury and determine whether metformin exerts its protective effect via miR-19a/ACSL axis. Firstly, bioinformatics analysis of data from Gene Expression Omnibus (GEO) database indicated that miR-19a was downregulated in patients with myocardial infarction (MI) compared to that in control group. H/R model was constructed with AC16 cells in vitro. qRT-PCR assay revealed that miR-19a was downregulated in H/R-treated AC16 cells. Then, CCK-8 assay demonstrated that upregulation of miR-19a significantly alleviated H/R-induced decline of cell viability. Moreover, bioinformatics prediction, western blotting and dual-luciferase reporter assays were performed to check the target genes of miR-19a, and ACSL1 was determined as a downstream target gene of miR-19a. Besides, the analysis based on Comparative Toxicogenomics Database (CTD) suggested that metformin targeting ACSL1 can be used as a potential drug for further research. Biological function experiments in vitro revealed that H/R markedly declined the viability and elevated the apoptosis of AC16 cells, while metformin can significantly mitigate these effects. Furthermore, overexpression of miR-19a significantly strengthened the beneficial effect of metformin on H/R-induced AC16 cells injury, which can be reversed by upregulation of ACSL1. In conclusion, metformin can alleviate H/R-induced cells injury via regulating miR-19a/ACSL axis, which lays a foundation for identifying novel targets for myocardial I/R injury therapy.
本研究旨在探讨 miR-19a/ACSL 轴在缺氧/复氧（H/R）诱导的心肌损伤中的作用，并确定二甲双胍是否通过 miR-19a/ACSL 轴发挥其保护作用。首先，通过基因表达综合数据库（GEO）数据库的生物信息学分析表明，与对照组相比，心肌梗死（MI）患者的 miR-19a 下调。体外采用 AC16 细胞构建 H/R 模型。qRT-PCR 检测显示 H/R 处理的 AC16 细胞中 miR-19a 下调。然后，CCK-8 检测表明 miR-19a 的上调显著减轻了 H/R 诱导的细胞活力下降。此外，通过生物信息学预测、Western blot 和双荧光素酶报告基因检测分析 miR-19a 的靶基因，确定 ACSL1 是 miR-19a 的下游靶基因。此外，基于比较毒理学基因组数据库（CTD）的分析表明，二甲双胍靶向 ACSL1 可作为进一步研究的潜在药物。体外生物学功能实验表明，H/R 显著降低了 AC16 细胞的活力并增加了其凋亡，而二甲双胍可显著减轻这些作用。此外，miR-19a 的过表达显著增强了二甲双胍对 H/R 诱导的 AC16 细胞损伤的有益作用，而 ACSL1 的上调可逆转这种作用。综上所述，二甲双胍通过调节 miR-19a/ACSL 轴减轻 H/R 诱导的细胞损伤，为识别心肌 I/R 损伤治疗的新靶点奠定了基础。
