Department of Cardiovascular Medicine, The Second Hospital of Hebei Medical University, No. 215 Heping West Road, Shijiazhuang, 050000, China.
Cell Stress Chaperones. 2023 Mar;28(2):177-190. doi: 10.1007/s12192-023-01328-x. Epub 2023 Feb 21.
Circular RNAs (circRNAs) have been identified as vital regulators in cardiovascular diseases, including acute myocardial infarction (AMI). In this study, the function and mechanism of circRNA heparan sulfate proteoglycan 2 (circHSPG2) in hypoxia-induced injury in AC16 cardiomyocytes were investigated. AC16 cells were stimulated with hypoxia to establish an AMI cell model in vitro. Real-time quantitative PCR and western blot assays were performed to quantify the expression levels of circHSPG2, microRNA-1184 (miR-1184), and mitogen-activated protein kinase kinase kinase 2 (MAP3K2). Counting Kit-8 (CCK-8) assay was used to measure cell viability. Flow cytometry was performed to detect cell cycle and apoptosis. Enzyme-linked immunosorbent assay (ELISA) was used to determine the expression of inflammatory factors. Dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays were used to analyze the relationship between miR-1184 and circHSPG2 or MAP3K2. In AMI serum, circHSPG2 and MAP3K2 mRNA were highly expressed and miR-1184 was down-regulated. Hypoxia treatment elevated HIF1α expression and repressed cell growth and glycolysis. Moreover, hypoxia promoted cell apoptosis, inflammation, and oxidative stress in AC16 cells. Hypoxia-induced circHSPG2 expression in AC16 cells. CircHSPG2 knockdown alleviated hypoxia-induced AC16 cell injury. CircHSPG2 directly targeted miR-1184, and miR-1184 targeted and suppressed MAP3K2. Inhibition of miR-1184 or overexpression of MAP3K2 abolished the alleviated effect of circHSPG2 knockdown on hypoxia-induced AC16 cell injury. Overexpression of miR-1184 relieved hypoxia-induced impairment in AC16 cells by MAP3K2. CircHSPG2 could regulate MAP3K2 expression through miR-1184. CircHSPG2 knockdown protected AC16 cells from hypoxia-induced injury by regulating the miR-1184/MAP3K2 cascade.
环状 RNA(circRNAs)已被确定为心血管疾病(包括急性心肌梗死(AMI))的重要调节因子。在这项研究中,研究了环状 RNA 硫酸乙酰肝素蛋白聚糖 2(circHSPG2)在体外缺氧诱导的 AC16 心肌细胞损伤中的功能和机制。用缺氧刺激 AC16 细胞建立 AMI 细胞模型。实时定量 PCR 和 Western blot 检测用于定量 circHSPG2、微小 RNA-1184(miR-1184)和丝裂原活化蛋白激酶激酶激酶 2(MAP3K2)的表达水平。使用细胞计数试剂盒-8(CCK-8)测定细胞活力。通过流式细胞术检测细胞周期和凋亡。酶联免疫吸附测定(ELISA)用于测定炎症因子的表达。双荧光素酶报告、RNA 免疫沉淀(RIP)和 RNA 下拉实验用于分析 miR-1184 与 circHSPG2 或 MAP3K2 的关系。在 AMI 血清中,circHSPG2 和 MAP3K2 mRNA 表达水平升高,miR-1184 表达水平下调。缺氧处理可升高 HIF1α 表达并抑制细胞生长和糖酵解。此外,缺氧可促进 AC16 细胞凋亡、炎症和氧化应激。AC16 细胞中 circHSPG2 的表达在缺氧诱导下增加。circHSPG2 敲低减轻了缺氧诱导的 AC16 细胞损伤。circHSPG2 直接靶向 miR-1184,miR-1184 靶向并抑制 MAP3K2。抑制 miR-1184 或过表达 MAP3K2 可消除 circHSPG2 敲低对缺氧诱导的 AC16 细胞损伤的缓解作用。miR-1184 的过表达通过 MAP3K2 缓解了缺氧对 AC16 细胞的损伤。circHSPG2 可以通过 miR-1184 调节 MAP3K2 的表达。circHSPG2 通过调节 miR-1184/MAP3K2 级联反应来保护 AC16 细胞免受缺氧诱导的损伤。