低氧预处理脐带间充质干细胞来源的外泌体对内皮细胞功能的影响

Effects of exosomes derived from hypoxia-preconditioned umbilical cord mesenchymal stem cells on function of endothelial cells

目的:探讨低氧预处理人脐带间充质干细胞(hUCMSCs)后,其释放的外泌体对人脐静脉内皮细胞(HUVECs)增殖、迁移和小管形成的影响。方法:分离、培养并鉴定hUCMSCs和HUVECs。采用超速离心法提取hUCMSCs释放的外泌体,透射电镜观察外泌体的形态,纳米颗粒跟踪分析技术检测外泌体的粒径和浓度,Western blot检测外泌体表面特异性标志蛋白。将hUCMSCs分为常氧组和低氧预处理组,用CCK-8法检测细胞活力。将HUVECs分为对照组、常氧外泌体组和低氧外泌体组,用EdU法检测细胞增殖能力,细胞划痕和Transwell实验检测细胞迁移能力,小管形成实验检测细胞小管形成能力。结果:与常氧组相比,低氧预处理增强hUCMSCs活力,促进其外泌体释放。与常氧外泌体组相比,低氧预处理hUCMSCs释放的外泌体增强HUVECs的增殖、迁移和小管形成能力。结论:低氧预处理hUCMSCs分泌的外泌体增强HUVECs增殖、迁移和小管形成能力。

AIM:To investigate the effect of exosomes derived from hypoxia-preconditioned human umbilical cord mesenchymal stem cells(hUCMSCs)on proliferation,migration and tube formation of human umbilical vein endothelial cells(HUVECs).METHODS:hUCMSCs and HUVECs were isolated,cultured and identified. Exosomes derived from hUCMSCs were extracted by ultracentrifugation. The morphological change of exosomes was observed under transmission electron microscope. The particle size and concentration of exosomes were detected by nanoparticle tracking analysis,and the surface specific marker proteins of exosomes were determined by Western blot. hUCMSCs were divided into normoxia group and hypoxia group. The viability of hUCMSCs was measured by CCK-8 assay. HUVECs were divided into control group,normoxic exosome group and hypoxic exosome group. The proliferation of HUVECs was detected by EdU assay. The migration ability was detected by cell scratch assay and Transwell experiment. Tube formation ability was evaluated by tube formation experiment.RESULTS:Compared with normoxia group,hypoxia pretreatment enhanced the viability and exosome release of hUCMSCs. Compared with normoxic exosome group,hypoxic exosomes enhanced the proliferation,migration and tube formation of HUVECs.CONCLUSION:Exosomes derived from hUCMSCs under hypoxia enhances the proliferation,migration and tube formation of HUVECs.