Extracellular vesicles from mesenchymal stem cells activates VEGF receptors and accelerates recovery of hindlimb ischemia
Extracellular vesicles from mesenchymal stem cells activates VEGF receptors and accelerates recovery of hindlimb ischemia
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DOI：
10.1016/j.jconrel.2017.08.022
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发表时间：
2017-10-28
影响因子：
10.8
通讯作者：
Ahn, Byeong-Cheol
中科院分区：
文献类型：
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作者：
Gangadaran, Prakash;Rajendran, Ramya Lakshmi;Ahn, Byeong-Cheol
来源于间充质干细胞（MSCs）的细胞外囊泡（EV）是治疗多种疾病的潜在药物，但其血管生成机制尚不清楚。在这里，我们描述了MSC-EV如何激活VEGF受体和下游血管生成途径。从细胞培养基中分离小鼠MSC-EV，并使用透射电子显微镜、纳米颗粒分析和蛋白质印迹进行表征。使用内皮细胞进行的体外迁移、增殖和管形成试验用于评估MSC-EV的血管生成潜力，并显示治疗后细胞迁移、增殖和管形成水平较高。qRT-PCR和Western blotting（WB）显示MSC-EV处理后小鼠SVEC-4内皮细胞中血管生成基因VEGFR 1和VEGFR 2的蛋白和mRNA表达增加。此外，其他重要的促血管生成途径（SRC、AKT和ERK）也被体外MSC-EV处理激活。WB和qRT-PCR显示MSC-EV中VEGF蛋白和miR-210- 3 p的富集存在。建立小鼠后肢缺血模型，将含或不含Matrigel的MSC-EVs（EV-MSC + Gel）注入缺血区，采用分子影像技术监测血液再灌注情况。MSC-EV的体内给药增加了缺血肢体中的血液再灌注和新血管的形成，加入基质胶通过缓慢释放EV进一步增强了这种效果。MSC-EV增强缺血肢体中的血管生成，最可能是通过内皮细胞中VEGFR 1和VEGFR 2的过表达。这些发现揭示了MSC-EV激活受体影响血管生成的新机制。
Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) are potential therapies for various diseases, but their angiogenic mechanisms of therapeutic efficacy remain unclear. Here, we describe how MSC-EVs, activates VEGF receptors and downstream angiogenesis pathways. Mouse MSC-EVs were isolated from cell culture medium and characterized using transmission electron microscopy, nanoparticle analysis, and western blotting. In vitro migration, proliferation, and tube formation assays using endothelial cells were used to assess the angiogenic potential of MSC-EVs, and revealed higher levels of cellular migration, proliferation, and tube formation after treatment. qRT-PCR and western blotting (WB) revealed higher protein and mRNA expression of the angiogenic genes VEGFR1 and VEGFR2 in mouse SVEC-4 endothelial cells after MSC-EVs treatment. Additionally, other vital pro-angiogenic pathways (SRC, AKT, and ERK) were activated by in vitro MSC-EV treatment. WB and qRT-PCR revealed enriched presence of VEGF protein and miR-210-3p in MSC-EV. The hindlimb ischemia mouse model was established and MSC-EVs with or without Matrigel (EV-MSC + Gel) were injected into the ischemic area and blood reperfusion was monitored using molecular imaging techniques. The in vivo administration of MSC-EVs increased both blood reperfusion and the formation of new blood vessels in the ischemic limb, with the addition of matrigel enhancing this effect further by releasing EVs slowly. MSC-EVs enhance angiogenesis in ischemic limbs, most likely via the overexpression of VEGFR1 and VEGFR2 in endothelial cells. These findings reveal a novel mechanism of activating receptors by MSC-EVs influence the angiogenesis.