Chen Xiao-Cui, Gai Min-Tao, He Chun-Hui, Zhao Bang-Hao, Liu Fen, Ma Xiang, Ma Yi-Tong, Gao Xiao-Ming, Chen Bang-Dang
State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, First Affiliated Hospital of Xinjiang Medical University, Clinical Medical Research Institute, Xinjiang Medical University, No. 137 Liyushan South Road, Urumqi, 830054, China.
College of Basic Medicine of Xinjiang Medical University, Urumqi, China.
Cardiovasc Drugs Ther. 2025 Jan 2. doi: 10.1007/s10557-024-07662-1.
To investigate the protective effect and mechanism of enhanced expression of endogenous macrophage migration inhibitory factor (MIF) on cardiac ischemia-reperfusion (I/R) injury.
A recombinant double-stranded adeno-associated virus serotype 9 with MIF or green fluorescent protein (GFP) genes (dsAAV9-MIF/GFP) was transduced into mice and neonatal rat ventricular myocytes (NRVMs). The models of cardiac 60 min ischemia and 24 h reperfusion and 12 h hypoxia/12 h reoxygenation (H/R) were established in mice and NRVMs, respectively. Infarct size, cardiac remodeling, and related signaling pathways were assessed.
The dsAAV9 vector demonstrated strong transduction efficacy and cardiac affinity. Cardiac overexpression of MIF led to a 35.3% reduction in infarct size and improved cardiac function following I/R injury. In the dsAAV9-MIF group, the AMP-activated protein kinase (AMPK) signaling pathway was activated, and autophagy was enhanced during the ischemic period. During reperfusion, the extracellular signal-regulated kinases 1 and 2 (ERK1/2) signaling pathway was upregulated, leading to reduced cardiac apoptosis. In vitro, transfection with MIF in NRVMs also upregulated AMPK and ERK1/2 signaling during hypoxia and reoxygenation, respectively. Furthermore, MIF overexpression significantly improved autophagy and mitochondrial function, evidenced by an increased LC3-II/I ratio and enhanced mitochondrial membrane potential (ΔΨm), with these effects reversed by the AMPK inhibitor compound C. Additionally, MIF overexpression led to a 60% reduction in the apoptosis rate of cardiomyocytes subjected to H/R and decreased the Bax/Bcl-2 ratio, partially through the ERK1/2 signaling pathway.
Enhanced endogenous MIF expression via the dsAAV9 vector provides significant cardioprotection against I/R injury by activating the AMPK and ERK1/2 signaling pathways. Our findings suggest that targeting MIF may represent a viable therapeutic strategy for severe and prolonged I/R injury.
研究内源性巨噬细胞移动抑制因子（MIF）表达增强对心脏缺血再灌注（I/R）损伤的保护作用及机制。
将携带MIF或绿色荧光蛋白（GFP）基因的重组9型双链腺相关病毒（dsAAV9-MIF/GFP）转导至小鼠和新生大鼠心室肌细胞（NRVMs）。分别在小鼠和NRVMs中建立心脏60分钟缺血及24小时再灌注模型和12小时缺氧/12小时复氧（H/R）模型。评估梗死面积、心脏重塑及相关信号通路。
dsAAV9载体显示出强大的转导效力和心脏亲和力。心脏MIF过表达导致I/R损伤后梗死面积减少35.3%，并改善心脏功能。在dsAAV9-MIF组中，缺血期AMP激活的蛋白激酶（AMPK）信号通路被激活，自噬增强。再灌注期间，细胞外信号调节激酶1和2（ERK1/2）信号通路上调，导致心脏细胞凋亡减少。在体外，NRVMs中MIF转染在缺氧和复氧期间也分别上调了AMPK和ERK1/2信号。此外，MIF过表达显著改善自噬和线粒体功能，表现为LC3-II/I比值增加和线粒体膜电位（ΔΨm）增强，而AMPK抑制剂化合物C可逆转这些作用。另外，MIF过表达使H/R处理的心肌细胞凋亡率降低60%，并降低Bax/Bcl-2比值，部分是通过ERK1/2信号通路实现的。
通过dsAAV9载体增强内源性MIF表达可通过激活AMPK和ERK1/2信号通路对I/R损伤提供显著的心脏保护作用。我们的研究结果表明，靶向MIF可能是治疗严重和长时间I/R损伤的可行治疗策略。
