缺血再灌注(I/R)是心血管疾病(CVD)和心脏移植的重要危险因素,I/R可引起心肌细胞缺氧/复氧(H/R)损伤。最近的研究表明,环状RNA(circularRNAs,circRNAs)可能影响H/R诱导的心肌损伤的进展,但机制仍然未知。我们的工作探讨了circ_0010729在H2O2诱导的心肌损伤中的作用。
通过定量实时聚合酶链反应(qRT-PCR)显示人心肌细胞(HCM)中circ_0010729,microRNA-1184(miR-1184)和受体相互作用丝氨酸/苏氨酸激酶1(RIPK1)的mRNA水平。同时,通过蛋白质印迹分析对RIPK1的蛋白水平进行定量.此外,通过5-乙炔基-29-脱氧尿苷(EdU)测定法检查细胞功能,流式细胞术测定,蛋白质印迹和抗氧化指标分析。此外,miR-1184和circ_0010729或RIPK1之间的相互作用通过双荧光素酶报告子试验检测.最终,体内实验用于测量circ_0010729的作用。
circ_0010729RNA和RIPK1蛋白水平升高,miR-1184在暴露于H2O2的HCM中降低。在功能分析中,circ_0010729缺乏抑制细胞凋亡和氧化应激,而在H2O2暴露下促进HCM细胞增殖。此外,miR-1184通过靶向RIPK1抑制H2O2诱导的心肌损伤。机械上,circ_0010729作为miR-1184海绵调控RIPK1的水平。
Circ_0010729促进H2O2诱导的心肌损伤,因此,circ_001729可以作为H/R诱导的心肌损伤的潜在疗法。
Ischemia-reperfusion (I/R) is an important risk factor for cardiovascular diseases (CVDs) and cardiac transplantation, as I/R can cause myocardial cell hypoxia/reoxygenation (H/R) injury. Recent research has shown that circular RNAs (circRNAs) may affect the progress of H/R-induced myocardial injury, but the mechanism remains unknown. Our work explored the role of
circ_0010729 in H2O2-induced myocardial injury.
The levels of circ_0010729, microRNA-1184 (miR-1184) and mRNA of receptor interacting serine/threonine kinase 1 (RIPK1) were indicated by quantitative real-time polymerase chain reaction (qRT-PCR) in human cardiac myocytes (HCMs). Meanwhile, the protein level of RIPK1 was quantified by western blot analysis. Besides, the cell functions were examined by 5-Ethynyl-29-deoxyuridine (EdU) assay, flow cytometry assay, western blot and antioxidant indexes analysis. Furthermore, the interplay between miR-1184 and
circ_0010729 or RIPK1 was detected by dual-luciferase reporter assay. Eventually, the in vivo experiments were applied to measure the role of
circ_0010729.
The levels of
circ_0010729 RNA and RIPK1 protein were increased, and the miR-1184 was decreased in HCMs exposed to H2O2. In functional analysis, circ_0010729 deficiency restrained cell apoptosis and oxidative stress, whereas promoted cell proliferation in HCMs under H2O2 exposure. Moreover, miR-1184 inhibited the H2O2-induced myocardial injury by targeting RIPK1. Mechanistically,
circ_0010729 acted as a miR-1184 sponge to regulate the level of RIPK1.
Circ_0010729 promotes H2O2-induced myocardial injury, and thus circ_001729 may be targeted as a potential therapy for H/R-induced myocardial injury.