Tsai Hsiao-Ya, Wang Jen-Chun, Hsu Yu-Juei, Lin Chih-Yuan, Huang Po-Hsun, Tsai Min-Chien, Hsu Chin-Wang, Yang Shang-Feng, Tsai Shih-Hung
Department of Emergency Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
Department of Emergency Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan; Institute of Clinical Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
Biomed J. 2024 Nov 23;48(4):100818. doi: 10.1016/j.bj.2024.100818.
Cardiac remodeling is implicated in numerous physiologic and pathologic conditions, including scar formation, heart failure, and cardiac arrhythmias. Nuclear factor-activated T-cell cytoplasmic (NFATc) is a crucial transcription factor that regulates cardiac remodeling. MicroRNA (miR)-424/322 has pathophysiological roles in the cardiovascular and respiratory systems by modulating hypoxia and inflammatory pathways. The role of miR-424/322 in regulating cardiac remodeling is under investigation. We identified several cardiac hypertrophy and fibrosis-related molecules as putative targets of miR-424/322. We propose that miR-424/322 could have crucial roles in cardiac remodeling by modulating several key molecules for cardiac fibrosis and hypertrophy.
Human cardiac fibroblasts (HCFs) and a myogenic cell line H9c2 cells were used for in vitro experiments. A murine model of angiotensin II (AngII)-induced cardiac remodeling was used to assess the roles of miR-322 on cardiac hypertrophy and fibrosis in vivo. Immunoblotting, immunofluorescence, real-time polymerase chain reaction and cell proliferation, Sirius Red, and dual-luciferase reporter assays were used to decipher the molecular mechanism.
We found that miR-322 knockout mice were susceptible to AngII-induced cardiac fibrosis and hypertrophy in vivo. Administration of miR-424/322 inhibitors aggravated AngII-induced overexpression of NFATc3, furin, natriuretic peptides and collagen 1A1 in H9c2 cells and HCFs. miR-424/322 mimics reversed the AngII-induced fibrosis, hypertrophy, and proliferation by targeting NFATc3 and furin in vitro. miR-424/322 could be transactivated by NFATc3. Exogenous miR-322 ameliorated AngII-induced hypertrophy and cardiac fibrosis in vivo.
The NFATc3/miR-424/322/furin axis is crucial for developing cardiac remodeling, and exogenous miR-322 mimics could have therapeutic potential in cardiac remodeling.
心脏重塑与多种生理和病理状况相关，包括瘢痕形成、心力衰竭和心律失常。核因子激活的T细胞胞质蛋白（NFATc）是调节心脏重塑的关键转录因子。微小RNA（miR）-424/322通过调节缺氧和炎症途径在心血管和呼吸系统中发挥病理生理作用。miR-424/322在调节心脏重塑中的作用正在研究中。我们确定了几种与心肌肥厚和纤维化相关的分子作为miR-424/322的假定靶点。我们提出，miR-424/322可能通过调节心肌纤维化和肥厚的几个关键分子在心脏重塑中发挥关键作用。
使用人心脏成纤维细胞（HCFs）和肌源性细胞系H9c2细胞进行体外实验。使用血管紧张素II（AngII）诱导的心脏重塑小鼠模型评估miR-322在体内对心肌肥厚和纤维化的作用。采用免疫印迹、免疫荧光、实时聚合酶链反应以及细胞增殖、天狼星红和双荧光素酶报告基因检测来阐明分子机制。
我们发现miR-322基因敲除小鼠在体内易受AngII诱导的心脏纤维化和肥厚影响。给予miR-424/322抑制剂会加重AngII诱导的H9c2细胞和HCFs中NFATc3、弗林蛋白酶、利钠肽和胶原蛋白1A1的过度表达。miR-424/322模拟物在体外通过靶向NFATc3和弗林蛋白酶逆转了AngII诱导的纤维化、肥厚和增殖。miR-424/322可被NFATc3反式激活。外源性miR-322改善了体内AngII诱导的肥厚和心脏纤维化。
NFATc3/miR-424/322/弗林蛋白酶轴对心脏重塑的发展至关重要，外源性miR-322模拟物在心脏重塑中可能具有治疗潜力。
