内皮祖细胞来源的外泌体减轻血管内皮细胞缺氧性损伤  

Endothelial Progenitor Cell Derived Exosomes Ameliorate Endothelial Cells Hypoxic Injury

作　　者：李罗成[1] 王志维[1] 吴红兵[1] 任宗力[1] 任伟[1] LI Luo-cheng;WANG Zhi-wei;WU Hong-bing;REN Zong-li;REN Wei(Department of Cardiovascular Surgery,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院心血管外科

出　　处：《微循环学杂志》2019年第4期1-6,共6页Chinese Journal of Microcirculation

基　　金：湖北省自然科学基金面上项目(2018CFB754)

摘　　要：目的:分析内皮祖细胞(EPCs)来源的外泌体(EXOs)对血管内皮细胞缺氧性损伤的影响。方法:体外培养EPCs,提取培养基中的EXOs并通过透射电镜和Western blot检测膜性标志物进行鉴定。将体外培养人脐静脉内皮细胞(HUVECs)分为3组:(1)正常对照组,常规培养的HUVECs;(2)缺氧组,将HUVECs于低氧环境中培养8h;(3)缺氧+EPCs-EXOs组,HUVECs于缺氧处理前加入EPCs来源的EXOs(EPCs-EXOs)处理,再在低氧环境中培养8h。于0h、24h和48h采用CCK-8增殖检测试剂盒、划痕实验及流式细胞术检测各组细胞的增殖、迁移和凋亡情况。结果:EPCs-EXOs在电镜下呈椭圆形或杯状膜性囊泡,直径约40-110nm,其膜性标志蛋白CD63、CD81呈阳性表达。与正常对照组比较,缺氧组细胞增殖和迁移能力下降,细胞凋亡率增加(P<0.05)。与缺氧组比较,缺氧+EPCs-EXOs组细胞增殖和迁移能力得到一定程度的恢复(P<0.01),与正常对照组相比无明显差异(P>0.05),同时其细胞凋亡率较缺氧组下降(P<0.01),但仍高于正常对照组(P<0.05)。结论:EPCs-EXOs可改善缺氧内皮细胞的增殖和迁移能力,减少内皮细胞凋亡,减轻血管内皮细胞的缺氧性损伤。Objective:To study the impact of endothelial progenitor cell-derived exosomes on hypoxic endothelial cell injury.Method:Endothelial progenitor cells were cultured in vitro and the culture medium was collected.Exosomes were isolated from the medium and identified with transmission electron microscope and western blot.Then in vitro cultured human umbilical vein endothelial cells(HUVECs)were divided into three groups:(1)Normal control group,HUVECs cultured in normal conditions.(2)Hypoxia group,HUVECs underwent 8 hours of hypoxia condition and then cultured in normal conditions.(3)Hypoxia+EPCs-EXOs group,HUVECs were treated with endothelial progenitor cell-derived exosomes before cultured in 8 hours of hypoxia condition.CCK-8/WST-8 kit,cell wound healing assay and flow cytometry were applied to evaluate the proliferation,migration and apoptosis of human umbilical vein endothelial cells.Results:Endothelial progenitor cell-derived cells derived exosomes were oval shaped vesicles with a diameter of 40-110 nm.The expression of CD63 and CD81 was positive.After being conditioned with hypoxia,human umbilical vein endothelial cells exhibited a compromised proliferation and migration capability,and an increased cell apoptosis(P<0.05).Endothelial progenitor cell-derived exosomes restored the proliferation and migration potential of human umbilical vein endothelial cells treated with hypoxia(P<0.01)and decreased endothelial cell apoptosis(P<0.01).Conclusion:Endothelial progenitor cell-derived exosomes enhance the proliferation and migration,and decrease the apoptosis of human umbilical vein endothelial cells treated with hypoxic condition,and thus ameliorate hypoxic endothelial cell injury.

关 键 词：内皮祖细胞 外泌体 内皮细胞 低氧 损伤 

分 类 号：R654[医药卫生—外科学]