Adipose-Derived Mesenchymal Stromal Cells Under Hypoxia: Changes in Extracellular Vesicles Secretion and Improvement of Renal Recovery after Ischemic Injury.
Adipose-Derived Mesenchymal Stromal Cells Under Hypoxia: Changes in Extracellular Vesicles Secretion and Improvement of Renal Recovery after Ischemic Injury.
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DOI：
10.33594/000000102
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发表时间：
2019-01-01
期刊：
影响因子：
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通讯作者：
Lindoso, Rafael Soares
中科院分区：
文献类型：
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作者：
Collino, Federica;Lopes, Jarlene Alecia;Lindoso, Rafael Soares
背景/目的：骨髓间充质干细胞(MSCs)来源的细胞外小泡(EVS)在肾脏损伤中的治疗潜力已有大量报道。然而，需要新的方法来优化肾脏疾病的治疗效果。MSCs生理上处于低氧分压(PO2)状态，在低氧条件下培养脂肪源性MSCs(ADMSCs)会改变其分泌旁分泌特性。本研究旨在探讨ADMSCs低氧预适应是否通过改变分泌EVS的特性来促进肾缺血再灌注损伤(IRI)后肾脏的恢复。方法：将培养的ADMSCs在21%pO2(对照组)和1%pO2(低氧)条件下培养上清液，超速离心分离EVS，并用流式细胞仪和电子显微镜对其进行鉴定。通过体外和体内模型分析这些EVS的摄取和作用。建立肾小管细胞株HK-2-DO-ATP耗竭损伤模型。用纳米UPLC串联纳米ESI-HDMSE方法对损伤后经EVS处理的细胞进行蛋白质组学分析。在活体分析中，雄性Wistar大鼠双侧缺血45min，然后在72小时再灌注期内肾囊内注射ADMSC-EVS。组织学、免疫组织化学和qRT-PCR分析这些肾脏的细胞死亡、炎症和氧化应激情况。结果：低氧可增加ADMSCs分泌EVS的能力，与常氧下分泌的EVS相比，ADMSCs分泌的EVS在肾组织的能量供应、抗细胞凋亡、免疫调节、血管生成和抗氧化应激反应方面存在差异。在常氧和低氧条件下，ADMSCs来源的EVS处理肾小管细胞的蛋白质组学分析给出了每种类型EVS的调节蛋白的特异性特征，表明了不同的生物学过程的调节。结论：综上所述，低氧可能为增强EVS在急性肾脏疾病治疗中的特性提供了一种有趣的策略。
BACKGROUND/AIMS: The therapeutic potential of extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) in kidney injury has been largely reported. However, new approaches are necessary to optimize the efficacy in the treatment of renal diseases. MSCs physiologically are under a low O2 partial pressure (pO2), and culturing adipose-derived MSCs (ADMSCs) in hypoxia alters their secretory paracrine properties. The aim of this study was to evaluate whether hypoxia preconditioning of ADMSCs alters the properties of secreted EVs to improve renal recovery after ischemia-reperfusion injury (IRI).METHODS: The supernatants of ADMSCs cultivated under 21% pO2 (control) or 1% pO2 (hypoxia) were ultracentrifuged for EVs isolation that were posteriorly characterized by flow cytometry and electron microscopy. The uptake and effects of these EVs were analyzed by using in vitro and in vivo models. HK-2 renal tubule cell line was submitted do ATP depletion injury model. Proteomic analyses of these cells treated with EVs after injury were performed by nano-UPLC tandem nano-ESI-HDMSE method. For in vivo analyses, male Wistar rats were submitted to 45 min bilateral ischemia, followed by renal intracapsular administration of ADMSC-EVs within a 72 h reperfusion period. Histological, immunohistochemical and qRT-PCR analysis of these kidneys were performed to evaluate cell death, inflammation and oxidative stress. Kidney function was evaluated by measuring the blood levels of creatinine and urea.RESULTS: The results demonstrate that hypoxia increases the ADMSCs capacity to secrete EVs that trigger different energy supply, antiapoptotic, immunomodulatory, angiogenic and anti-oxidative stress responses in renal tissue compared with EVs secreted in normoxia. Proteomic analyses of renal tubule cells treated with EVs from ADMSCs in normoxia and hypoxia give a specific signature of modulated proteins for each type of EVs, indicating regulation of distinct biological processes.CONCLUSION: In summary, hypoxia potentially offers an interesting strategy to enhance the properties of EVs in the treatment of acute kidney disease.