Surface Phosphatidylserine Is Responsible for the Internalization on Microvesicles Derived from Hypoxia-Induced Human Bone Marrow Mesenchymal Stem Cells into Human Endothelial Cells.
Surface Phosphatidylserine Is Responsible for the Internalization on Microvesicles Derived from Hypoxia-Induced Human Bone Marrow Mesenchymal Stem Cells into Human Endothelial Cells.
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表面磷脂酰丝氨酸负责缺氧诱导的人骨髓间充质干细胞衍生的微泡进入人内皮细胞
DOI：
10.1371/journal.pone.0147360
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Zhang H
中科院分区：
文献类型：
--
作者：
Wei X;Liu C;Wang H;Wang L;Xiao F;Guo Z;Zhang H
研究背景低氧诱导的间充质干细胞（MSC-MVs）微囊泡可被内皮细胞内化，促进内皮细胞增殖和血管结构形成，促进体内血管生成。然而，关于MSC-MV如何被内皮细胞摄取的信息很少。方法将人骨髓间充质干细胞在缺氧和/或无血清条件下培养，取上清液制备微血管。流式细胞术和共聚焦显微镜观察在存在或不存在重组人Annexin-V（Anx-V）和抗人CD 29和CD 44抗体的情况下，缺氧诱导的MSC-MV掺入人脐带内皮细胞（HUVECs）。此外，将靶向Anx-V和PSR的小干扰RNA（siRNA）递送到HUVEC中，或者用针对磷脂酰丝氨酸受体（PSR）的单克隆抗体处理HUVEC，并重新评估MV的细胞内化。结果外源性Anx-V可抑制HUVECs对缺氧诱导干细胞MVs的摄取，且呈剂量和时间依赖性，而抗CD 29和CD 44抗体对MVs的内化过程无影响。在Anx-V siRNA转染的HUVECs中也没有观察到抑制，然而，添加抗PSR抗体和PSR siRNA转染的HUVECs极大地阻断了从缺氧诱导的干细胞分离的MV掺入HUVECs。结论缺氧诱导的干细胞分离的微血管上的PS是HUVECs摄取PS的关键分子。
Background Previous data have proven that microvesicles derived from hypoxia-induced mesenchymal stem cells (MSC-MVs) can be internalized into endothelial cells, enhancing their proliferation and vessel structure formation and promoting in vivo angiogenesis. However, there is a paucity of information about how the MSC-MVs are up-taken by endothelial cells. Methods MVs were prepared from the supernatants of human bone marrow MSCs that had been exposed to a hypoxic and/or serum-deprivation condition. The incorporation of hypoxia-induced MSC-MVs into human umbilical cord endothelial cells (HUVECs) was observed by flow cytometry and confocal microscopy in the presence or absence of recombinant human Annexin-V (Anx-V) and antibodies against human CD29 and CD44. Further, small interfering RNA (siRNA) targeted at Anx-V and PSR was delivered into HUVECs, or HUVECs were treated with a monoclonal antibody against phosphatidylserine receptor (PSR) and the cellular internalization of MVs was re-assessed. Results The addition of exogenous Anx-V could inhibit the uptake of MVs isolated from hypoxia-induced stem cells by HUVECs in a dose- and time-dependent manner, while the anti-CD29 and CD44 antibodies had no effect on the internalization process. The suppression was neither observed in Anx-V siRNA-transfected HUVECs, however, addition of anti-PSR antibody and PSR siRNA-transfected HUVECs greatly blocked the incorporation of MVs isolated from hypoxia-induced stem cells into HUVECs. Conclusion PS on the MVs isolated from hypoxia-induced stem cells is the critical molecule in the uptake by HUVECs.