Wang Feng-Dan, Ding Yi, Zhou Jian-Hong, Zhou En, Zhang Tian-Tian, Fan Yu-Qi, He Qing, Zhang Zong-Qi, Mao Cheng-Yu, Zhang Jun-Feng, Zhou Jing
Department of Cardiology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.
Department of Anesthesiology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.
World J Stem Cells. 2024 Oct 26;16(10):873-895. doi: 10.4252/wjsc.v16.i10.873.
Myocardial ischemia-reperfusion injury (MIRI) poses a prevalent challenge in current reperfusion therapies, with an absence of efficacious interventions to address the underlying causes.
To investigate whether the extracellular vesicles (EVs) secreted by adipose mesenchymal stem cells (ADSCs) derived from subcutaneous inguinal adipose tissue (IAT) under γ-aminobutyric acid (GABA) induction (GABA-EVs) demonstrate a more pronounced inhibitory effect on mitochondrial oxidative stress and elucidate the underlying mechanisms.
We investigated the potential protective effects of EVs derived from mouse ADSCs pretreated with GABA. We assessed cardiomyocyte injury using terminal deoxynucleotidyl transferase dUTP nick end-labeling and Annexin V/propidium iodide assays. The integrity of cardiomyocyte mitochondria morphology was assessed using electron microscopy across various intervention backgrounds. To explore the functional RNA diversity between EVs and GABA-EVs, we employed microRNA (miR) sequencing. Through a dual-luciferase reporter assay, we confirmed the molecular mechanism by which EVs mediate thioredoxin-interacting protein (TXNIP). Western blotting and immunofluorescence were conducted to determine how TXNIP is involved in mediation of oxidative stress and mitochondrial dysfunction.
Our study demonstrates that, under the influence of GABA, ADSCs exhibit an increased capacity to encapsulate a higher abundance of miR-21-5p within EVs. Consequently, this leads to a more pronounced inhibitory effect on mitochondrial oxidative stress compared to EVs from ADSCs without GABA intervention, ultimately resulting in myocardial protection. On a molecular mechanism level, EVs regulate the expression of TXNIP and mitigating excessive oxidative stress in mitochondria during MIRI process to rescue cardiomyocytes.
Administration of GABA leads to the specific loading of miR-21-5p into EVs by ADSCs, thereby regulating the expression of TXNIP. The EVs derived from ADSCs treated with GABA effectively ameliorates mitochondrial oxidative stress and mitigates cardiomyocytes damage in the pathological process of MIRI.
心肌缺血再灌注损伤（MIRI）是当前再灌注治疗中普遍存在的挑战，缺乏有效的干预措施来解决其根本原因。
研究γ-氨基丁酸（GABA）诱导下腹股沟皮下脂肪组织（IAT）来源的脂肪间充质干细胞（ADSCs）分泌的细胞外囊泡（EVs）（GABA-EVs）是否对线粒体氧化应激具有更显著的抑制作用，并阐明其潜在机制。
我们研究了经GABA预处理的小鼠ADSCs来源的EVs的潜在保护作用。使用末端脱氧核苷酸转移酶dUTP缺口末端标记法和膜联蛋白V/碘化丙啶检测法评估心肌细胞损伤。在不同干预背景下，通过电子显微镜评估心肌细胞线粒体形态的完整性。为了探索EVs和GABA-EVs之间的功能性RNA多样性，我们采用了微小RNA（miR）测序。通过双荧光素酶报告基因检测，我们证实了EVs介导硫氧还蛋白相互作用蛋白（TXNIP）的分子机制。进行蛋白质免疫印迹和免疫荧光检测，以确定TXNIP如何参与氧化应激和线粒体功能障碍的介导。
我们的研究表明，在GABA的影响下，ADSCs在EVs中封装更高丰度miR-21-5p的能力增强。因此，与未经过GABA干预的ADSCs来源的EVs相比，这对线粒体氧化应激具有更显著的抑制作用，最终实现心肌保护。在分子机制层面，EVs在MIRI过程中调节TXNIP的表达并减轻线粒体中的过度氧化应激，从而挽救心肌细胞。
给予GABA可导致ADSCs将miR-21-5p特异性加载到EVs中，从而调节TXNIP的表达。GABA处理的ADSCs来源的EVs在MIRI的病理过程中有效改善线粒体氧化应激并减轻心肌细胞损伤。
