Yu Ziyang, Xu Wenbo, Teng Yirong, Li Tingting, Guo Ren, Li Ju, Li Xichen, Li Yanping, Hao Yinglu
Department of Cardiology, The 6th Affiliated Hospital of Kunming Medical University, The People's Hospital of Yuxi City.
Department of Laboratory, The 6th Affiliated Hospital of Kunming Medical University, The People's Hospital of Yuxi City.
Eur J Histochem. 2025 Jun 17;69(3). doi: 10.4081/ejh.2025.4230. Epub 2025 Aug 22.
Cardiomyocyte injury related to hypoxia/reoxygenation (H/R) is pivotal in myocardial infarction. The circular RNA circRNA-79530 (circ79530) may play a regulatory role in this process, though its exact function has yet to be elucidated. This research explores the role of circRNA-79530 in H9c2 cells under H/R, with a particular focus on its interactions with miR-214 and the transcription factor Twist. It also examines their subsequent effects on mitochondrial function and oxidative stress. H9c2 cardiomyocytes were subjected to H/R to model myocardial injury. We measured circRNA-79530, miR-214, and Twist levels via RT-qPCR, with Twist protein via Western blotting. ROS levels were quantified using DCFH-DA, and cell viability and injuries were assessed through CCK-8, LDH, SOD, and MDA assays, respectively. Mitochondrial performance was assessed through various methods, including the measurement of mitochondrial membrane potential using JC-1 staining, the quantification of ATP levels, and the examination of the protein levels of mitochondrial complexes, as well as the expression of fusion proteins. Our findings indicated that downregulation of circRNA-79530 modulated miR-214 and Twist expression, influencing mitochondrial dynamics and ROS production. Knockdown of circRNA-79530 improved cell viability, reduced oxidative stress and enhanced mitochondrial function. Additionally, overexpression of miR-214 mitigated Twist expression, further supporting the effect of miR-214 in H/R conditions. circRNA-79530 could worsen oxidative stress and mitochondrial dysfunction, and regulate Twist-mediated mitochondrial damage via sponging miR-214 in H9c2 cells under H/R conditions.
与缺氧/复氧（H/R）相关的心肌细胞损伤在心肌梗死中起关键作用。环状RNA circRNA - 79530（circ79530）可能在此过程中发挥调节作用，但其确切功能尚待阐明。本研究探讨circRNA - 79530在H/R条件下对H9c2细胞的作用，特别关注其与miR - 214和转录因子Twist的相互作用。还研究了它们随后对线粒体功能和氧化应激的影响。对H9c2心肌细胞进行H/R处理以模拟心肌损伤。通过RT-qPCR检测circRNA - 79530、miR - 214和Twist水平，通过蛋白质免疫印迹检测Twist蛋白。使用DCFH-DA定量ROS水平，分别通过CCK-8、LDH、SOD和MDA检测评估细胞活力和损伤。通过多种方法评估线粒体性能，包括使用JC-1染色测量线粒体膜电位、定量ATP水平、检测线粒体复合物的蛋白质水平以及融合蛋白的表达。我们的研究结果表明，circRNA - 79530的下调调节了miR - 214和Twist的表达，影响线粒体动力学和ROS产生。敲低circRNA - 79530可提高细胞活力，降低氧化应激并增强线粒体功能。此外，miR - 214的过表达减轻了Twist的表达，进一步支持了miR - 214在H/R条件下的作用。在H/R条件下，circRNA - 79530可加重氧化应激和线粒体功能障碍，并通过在H9c2细胞中吸附miR - 214来调节Twist介导的线粒体损伤。
