Circ_0010729 regulates hypoxia-induced cardiomyocyte injuries by activating TRAF5 via sponging miR-27a-3p
Circ_0010729 regulates hypoxia-induced cardiomyocyte injuries by activating TRAF5 via sponging miR-27a-3p
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DOI：
10.1016/j.lfs.2020.118511
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发表时间：
2020-12-01
期刊：
影响因子：
6.1
通讯作者：
Wang, Zhifang
中科院分区：
文献类型：
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作者：
Lei, Dazhou;Wang, Yan;Wang, Zhifang
缺血性心肌病是一种严重的心血管疾病，死亡率高。环状RNA（circRNA）在包括缺血性心肌病在内的多种人类疾病中受到广泛调控。本研究旨在探讨circRNA circ_0010729在缺氧诱导的心肌细胞损伤中的作用机制。将人心肌细胞（AC 16）暴露于缺氧以模拟体外缺血性心肌病。分别采用MTT法、流式细胞术和糖酵解应激试验检测细胞活力、凋亡/坏死和糖酵解进程。细胞凋亡也通过切割的半胱天冬酶-3/7的活性来评估。免疫印迹法检测糖酵解相关蛋白和肿瘤坏死因子受体相关因子5（TRAF5）的表达。通过实时定量逆转录-聚合酶链反应（qRT-PCR）测量circ_0010729和miR-27 a-3p的表达。通过双荧光素酶报告基因实验、RNA免疫沉淀（RIP）实验和RNA pull-down实验对靶向关系的预测进行了验证。结果表明，低氧处理抑制细胞活力，诱导细胞凋亡和阻断糖酵解，然而，这些损伤被circ_0010729敲低减轻。miR-27a-3p被circ_0010729靶向，miR-27a-3p抑制逆转了circ_0010729敲低的作用，导致细胞损伤恶化。此外，TRAF5是miR-27a-3p的靶标，并且circ_0010729通过海绵状吸收miR-27a-3p上调TRAF5的表达。miR-27 a-3p的恢复增强了细胞活力，耗尽了细胞凋亡，促进了缺氧诱导的AC16细胞的糖酵解，而TRAF5过表达则消除了这些作用。总之，circ_0010729敲低通过介导miR-27 a-3p/TRAF5轴减轻缺氧诱导的AC16细胞损伤。
Ischemic cardiomyopathy is a severe cardiovascular disease with high mortality. Circular RNAs (circRNAs) are widely regulated in diverse human diseases, including Ischemic cardiomyopathy. This study aimed to investigate a novel functional mechanism of circRNA circ_0010729 in hypoxia-induced cardiomyocyte injuries. Human cardiomyocytes (AC16) were exposed to hypoxia to mimic ischemic cardiomyopathy in vitro. Cell viability, apoptosis/necrosis and glycolysis progress, were determined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, flow cytometry assay and glycolysis stress test, respectively. Cell apoptosis was also assessed by the activity of cleaved caspase-3/7. The levels of glycolysis-related proteins and tumor necrosis factor receptor-associated factor 5 (TRAF5) were examined by western blot. The expression of circ_0010729 and miR-27a-3p was measured by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The prediction about the targeted relationship was verified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. As a result, hypoxia treatment inhibited cell viability, induced cell apoptosis and blocked glycolysis, however, these injuries were alleviated by circ_0010729 knockdown. MiR-27a-3p was targeted by circ_0010729, and miR-27a-3p inhibition reversed the role of circ_0010729 knockdown, leading to the deterioration of cell injuries. Further, TRAF5 was a target of miR-27a-3p, and circ_0010729 upregulated the expression of TRAF5 by sponging miR-27a-3p. MiR-27a-3p restoration enhanced cell viability, depleted cell apoptosis and promoted glycolysis of hypoxia-induced AC16 cells, while these effects were abolished by TRAF5 overexpression. In conclusion, circ_0010729 knockdown alleviated hypoxia-induced AC16 cell injuries by mediating the miR-27a-3p/TRAF5 axis.