Lai Tsai-Chun, Lee Tzu-Lin, Chang Yu-Chun, Chen Yu-Chen, Lin Shu-Rung, Lin Shu-Wha, Pu Chi-Ming, Tsai Jaw-Shiun, Chen Yuh-Lien
Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
Department of Bioscience Technology, College of Science, Chung Yuan Christian University, Taoyuan, Taiwan.
Front Cell Dev Biol. 2020 Dec 3;8:569150. doi: 10.3389/fcell.2020.569150. eCollection 2020.
Cardiovascular disease is a major health problem in industrialized and developing countries and is the leading cause of death and disability. Myocardial ischemia/reperfusion (I/R) causes cardiomyocyte damage such as apoptosis and hypertrophy. The purpose of this study was to investigate the effects of exosomes from adipose-derived stem cells (ADSC-Exo) on hearts from I/R mice and to explore the underlying mechanisms. ADSC-Exo significantly decreased I/R-induced cardiomyocyte apoptosis and hypertrophy, as detected by TdT-mediated dUTP nick end-labeling (TUNEL) and wheat germ agglutinin (WGA) staining, respectively. In addition, the expression of apoptosis-related proteins p-p53 and PUMA and hypertrophy-related proteins ETS-1 and ANP were significantly reduced in the cardiomyocytes of ADSC-Exo-treated I/R mice compared to those of control mice. Both PUMA and ETS-1 are reported to be target genes for miR-221/222. I/R operation significantly reduced miR-221/222 expression, while ADSC-Exo treatment increased miR-221/222 expression, as detected by RT-qPCR. We also observed that cardiac I/R operation markedly increased cell apoptosis and hypertrophy in miR-221/222 knockout (KO) mice, while ADSC-Exo reduced the effects of I/R operation. Furthermore, ADSC-Exo protected H9c2 cardiomyocytes from HO-induced damage by reducing apoptosis and hypertrophy . HO treatment significantly reduced miR-221/222 expression, while ADSC-Exo treatment reversed this effect in H9c2 cells. ADSC-Exo treatment decreased HO-induced PUMA and ETS-1 expression. Compared with control treatment, I/R treatment significantly reduced p-AKT and increased p-p65, while ADSC-Exo and miR-221/222 mimics attenuated these effects. The AKT activator SC79 and p65 inhibitor Bay 11-7082 reduced HO-induced cell apoptosis and hypertrophy. Based on these findings, ADSC-Exo prevents cardiac I/R injury through the miR-221/miR-222/PUMA/ETS-1 pathway. Therefore, ADSC-Exo is an effective inhibitor of I/R-induced heart injury.
心血管疾病在工业化国家和发展中国家都是一个主要的健康问题，并且是死亡和残疾的主要原因。心肌缺血/再灌注（I/R）会导致心肌细胞损伤，如凋亡和肥大。本研究的目的是探讨脂肪来源干细胞外泌体（ADSC-Exo）对I/R小鼠心脏的影响，并探究其潜在机制。分别通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）和麦胚凝集素（WGA）染色检测发现，ADSC-Exo显著降低了I/R诱导的心肌细胞凋亡和肥大。此外，与对照小鼠相比，ADSC-Exo处理的I/R小鼠心肌细胞中凋亡相关蛋白p-p53和PUMA以及肥大相关蛋白ETS-1和心钠素（ANP）的表达显著降低。据报道，PUMA和ETS-1都是miR-221/222的靶基因。通过逆转录定量聚合酶链反应（RT-qPCR）检测发现，I/R手术显著降低了miR-221/222的表达，而ADSC-Exo处理则增加了miR-221/222的表达。我们还观察到，在miR-221/222基因敲除（KO）小鼠中，心脏I/R手术显著增加了细胞凋亡和肥大，而ADSC-Exo减轻了I/R手术的影响。此外，ADSC-Exo通过减少凋亡和肥大保护H9c2心肌细胞免受过氧化氢（HO）诱导的损伤。HO处理显著降低了miR-221/222的表达，而ADSC-Exo处理在H9c2细胞中逆转了这种作用。ADSC-Exo处理降低了HO诱导的PUMA和ETS-1表达。与对照处理相比，I/R处理显著降低了磷酸化蛋白激酶B（p-AKT）水平并增加了磷酸化核因子κB（p-p65）水平，而ADSC-Exo和miR-221/222模拟物减弱了这些作用。AKT激活剂SC79和p65抑制剂Bay 11-7082减少了HO诱导的细胞凋亡和肥大。基于这些发现，ADSC-Exo通过miR-221/miR-222/PUMA/ETS-1途径预防心脏I/R损伤。因此，ADSC-Exo是I/R诱导的心脏损伤的有效抑制剂。
