Department of Cardiology, Subei People's Hospital, School of Medicine, Yangzhou University, 98 Nantong West Road, Yangzhou, 225001, People's Republic of China.
Department of Critical Care Medicine, Subei People's Hospital, School of Medicine, Yangzhou University, 98 Nantong West Road, Yangzhou, 225001, People's Republic of China.
Stem Cell Res Ther. 2017 Sep 29;8(1):211. doi: 10.1186/s13287-017-0662-7.
Mesenchymal stem cells microvesicles (MSC-MVs) stabilize endothelial barrier function in acute lung injury (ALI); however, the detailed mechanism remains to be further defined. Hepatocyte growth factor (HGF), which is derived from MSC-MVs, might have a key role in the restoration of endothelial barrier function by MSC-MVs.
MSCs with lentiviral vector-mediated HGF gene knockdown (siHGF-MSC) were generated. A co-culture model of pulmonary microvascular endothelial cells and MSC-MVs collected from MSCs or siHGF-MSCs after 24 h of hypoxic culture was utilized. Then, endothelial paracellular and transcellular permeabilities were detected. VE-cadherin, and occludin protein expression in the endothelial cells was measured using Western blot. Endothelial cell proliferation was analysed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. Endothelial cell apoptosis was analysed using TUNEL assay. Finally, IL-6 and IL-10 production was determined via an enzyme-linked immunosorbent assay (ELISA).
Treatment with MSC-MVs significantly decreased LPS-induced endothelial paracellular and transcellular permeabilities, and the effect was significantly inhibited after HGF gene knockdown in MSC-MVs. Furthermore, treatment with MSC-MVs increased the expression of the endothelial intercellular junction proteins VE-cadherin and occludin. Treatment with MSC-MVs also decreased endothelial apoptosis and induced endothelial cell proliferation. Finally, the treatment reduced IL-6 production and increased IL-10 production in the conditioned media of endothelial cells. However, the effects of the treatment with MSC-MVs were inhibited after HGF gene knockdown.
MSC-MVs protect the barrier functions of pulmonary microvascular endothelial cells, which can be partly attributed to the presence of HGF in the MSC-MVs.
间充质干细胞微囊泡（MSC-MVs）可稳定急性肺损伤（ALI）中的内皮屏障功能；然而，其详细机制仍有待进一步确定。来源于 MSC-MVs 的肝细胞生长因子（HGF）可能在 MSC-MVs 恢复内皮屏障功能中起关键作用。
利用慢病毒载体介导的 HGF 基因敲低（siHGF-MSC）生成 MSC。建立肺微血管内皮细胞与 MSC 或 siHGF-MSC 缺氧培养 24 小时后收集的 MSC-MVs 的共培养模型。然后检测内皮细胞旁通透性和跨细胞通透性。采用 Western blot 法检测内皮细胞 VE-钙黏蛋白和闭合蛋白的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑（MTT）试验分析内皮细胞增殖。采用 TUNEL 试验分析内皮细胞凋亡。最后，通过酶联免疫吸附试验（ELISA）测定 IL-6 和 IL-10 的产生。
MSC-MVs 处理可显著降低 LPS 诱导的内皮细胞旁通透性和跨细胞通透性，而在 MSC-MVs 中敲低 HGF 基因后，这种作用明显受到抑制。此外，MSC-MVs 处理可增加内皮细胞间连接蛋白 VE-钙黏蛋白和闭合蛋白的表达。MSC-MVs 处理还可减少内皮细胞凋亡并诱导内皮细胞增殖。最后，该处理减少了内皮细胞条件培养基中 IL-6 的产生，增加了 IL-10 的产生。然而，在 MSC-MVs 中敲低 HGF 基因后，该处理的作用受到抑制。
MSC-MVs 可保护肺微血管内皮细胞的屏障功能，这在一定程度上归因于 MSC-MVs 中存在的 HGF。
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