Yuan Fei, Liu Jie, Zhong Liang, Liu Pengtao, Li Ting, Yang Kexin, Gao Wei, Zhang Guangyuan, Sun Jie, Zou Xiangyu
Department of Urology, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China.
Department of Neurology, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Stem Cell Res Ther. 2025 Feb 4;16(1):39. doi: 10.1186/s13287-025-04166-z.
Extracellular vesicles (EVs) secreted by mesenchymal stromal cells (MSCs) have been shown to provide significant protection against renal ischemia-reperfusion injury (IRI). Hypoxia has emerged as a promising strategy to enhance the tissue repair capabilities of MSCs. However, the specific effects of hypoxia on MSCs and MSC-EVs, as well as their therapeutic potential in renal IRI, remain unclear. In this study, we investigated the alterations occurring in MSCs and the production of MSC-EVs following hypoxia pre-treatment, and further explored the key intrinsic mechanisms underlying the therapeutic effects of hypoxic MSC-EVs in the treatment of renal IRI.
Human umbilical cord MSCs were cultured under normoxic and hypoxic conditions. Proliferation and related pathways were measured, and RNA sequencing was used to detect changes in the transcriptional profile. MSC-EVs from both normoxic and hypoxic conditions were isolated and characterized. In vivo, the localization and therapeutic effects of MSC-EVs were assessed in a rat renal IRI model. Histological examinations were conducted to evaluate the structure, proliferation, and apoptosis of IRI kidney tissue respectively. Renal function was assessed by measuring serum creatinine and blood urea nitrogen levels. In vitro, the therapeutic potential of MSC-EVs were measured in renal tubular epithelial cells injured by antimycin A. Protein sequencing analysis of hypoxic MSC-EVs was performed, and the depletion of Glutathione S-Transferase Omega 1 (GSTO1) in hypoxic MSC-EVs was carried out to verify its key role in alleviating renal injury.
Hypoxia alters MSCs transcriptional profile, promotes their proliferation, and increases the production of EVs. Hypoxia-pretreated MSC-EVs demonstrated a superior ability to mitigate renal IRI, enhancing proliferation and reducing apoptosis of renal tubular epithelial cells both in vivo and in vitro. Protein profiling of the EVs revealed an accumulation of numerous anti-oxidative stress proteins, with GSTO1 being particularly prominent. Knockdown of GSTO1 significantly reduced the antioxidant and therapeutic effects on renal IRI of hypoxic MSC-EVs.
Hypoxia significantly promotes the generation of MSC-EVs and enhances their therapeutic effects on renal IRI. The antioxidant stress effect induced by GSTO1 is identified as one of the most critical underlying mechanisms. Our findings highlight that hypoxia-pretreated MSC-EVs represent a novel and promising therapeutic strategy for renal IRI.
间充质基质细胞（MSC）分泌的细胞外囊泡（EV）已被证明能为肾缺血再灌注损伤（IRI）提供显著保护。缺氧已成为增强MSC组织修复能力的一种有前景的策略。然而，缺氧对MSC和MSC-EV的具体影响及其在肾IRI中的治疗潜力仍不清楚。在本研究中，我们调查了缺氧预处理后MSC发生的变化以及MSC-EV的产生，并进一步探索了缺氧MSC-EV治疗肾IRI的关键内在机制。
人脐带MSC在常氧和缺氧条件下培养。测量增殖及相关通路，并使用RNA测序检测转录谱的变化。分离并鉴定常氧和缺氧条件下的MSC-EV。在体内，在大鼠肾IRI模型中评估MSC-EV的定位和治疗效果。进行组织学检查以分别评估IRI肾组织的结构、增殖和凋亡。通过测量血清肌酐和血尿素氮水平评估肾功能。在体外，在抗霉素A损伤的肾小管上皮细胞中测量MSC-EV的治疗潜力。对缺氧MSC-EV进行蛋白质测序分析，并对缺氧MSC-EV中的谷胱甘肽S-转移酶ω1（GSTO1）进行敲低以验证其在减轻肾损伤中的关键作用。
缺氧改变MSC转录谱，促进其增殖，并增加EV的产生。缺氧预处理的MSC-EV表现出更强的减轻肾IRI的能力，在体内和体外均能增强肾小管上皮细胞的增殖并减少其凋亡。EV的蛋白质谱分析显示大量抗氧化应激蛋白积累，其中GSTO1尤为突出。敲低GSTO1显著降低了缺氧MSC-EV对肾IRI的抗氧化和治疗效果。
缺氧显著促进MSC-EV的生成并增强其对肾IRI的治疗效果。由GSTO1诱导的抗氧化应激效应被确定为最关键的潜在机制之一。我们的研究结果表明，缺氧预处理的MSC-EV代表了一种用于肾IRI的新型且有前景的治疗策略。
