人胎盘间充质干细胞外泌体促进人肺血管内皮细胞血管形成和迁移  

Exosomes from Human Placental Mesenchymal Stem Cell Promote Angiogenesis and Migration of Human Pulmonary Vascular Endothelial Cells

作　　者：吕玉珍 喻文琴 马晓薇 LYU Yuzhen;YU Wenqin;MA Xiaowei(Ningxia Medical University,Yinchuan 750004,China;Human Stem Cell Institute of General Hospital of Ningxia Medical University,Yinchuan 750004,China;Intensive Care Unit,Cardiocerebral Vascular Disease Hospital,General Hospital of Ningxia Medical University,Yinchuan 750012,China)

机构地区：[1]宁夏医科大学,银川750004 [2]宁夏医科大学总医院干细胞研究所,银川750004 [3]宁夏医科大学总医院心脑血管病医院ICU,银川750012

出　　处：《宁夏医科大学学报》2022年第7期649-655,662,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81860350);宁夏自然科学基金项目(2020AAC02002)。

摘　　要：目的探讨人胎盘间充质干细胞外泌体(HPMSCs-EX)对脂多糖(LPS)诱导损伤的人肺血管内皮细胞(HPVECs)血管形成和迁移能力的影响。方法采用酶消化法提取人胎盘间充质干细胞(HPMSCs),显微镜下观察细胞形态及诱导分化能力;从培养上清中分离纯化HPMSCs-EX,对其生物学特征进行鉴定,并利用荧光显微镜观察HPVECs对HPMSCs-EX摄取情况。将HPVECs分为对照组(NC组)、外泌体组(NC+HPMSCs-EX组)、损伤组(LPS组)及外泌体治疗组(LPS+HPMSCs-EX组),通过血管形成实验、迁移实验、划痕实验、免疫荧光实验及Western blot实验检测HPMSCs-EX对HPVECs血管形成、迁移能力的影响。结果显微镜下显示HPMSCs形似柳叶状,整体呈漩涡状生长,具备成脂肪、成骨及成软骨分化能力;透射电镜下显示HPMSCs-EX呈圆饼状,直径<150 nm;Western blot实验显示HPMSCs-EX表面特异抗原CD9、CD63表达呈阳性,Calnexin表达呈阴性;HPMSCs-EX能被HPVECs摄取。NC组和NC+HPMSCs-EX组实验结果差异无统计学意义(P均>0.05);LPS组细胞形态改变,细胞间连接松散,血管形成和迁移能力下降(P均<0.05),VEGFA和Vimentin表达下调(P均<0.05);LPS+HPMSCs-EX组细胞普遍增大,部分细胞变圆、变亮飘起,但多数细胞形态尚存,细胞血管形成和迁移能力改善(P均<0.05),VEGFA和Vimentin表达上调(P均<0.05)。结论HPMSCs-EX可能通过促进血管形成与迁移能力改善LPS诱导损伤的HPVECs功能。Objective To investigate the effects of human placenta mesenchymal stem cell exosomes(HPMSCs-EX)on angiogenesis and migration of human pulmonary vascular endothelial cells(HPVECs)injured by lipopolysaccharide(LPS).Methods HPMSCs were extracted by enzymatic digestion,and the cell morphology and differentiation ability were observed under microscope.HPMSCs-EX were isolated and purified from the culture supernatant,and its biological characteristics were identified.The uptake of HPMSCs-EX by HPVECs was observed by fluorescence microscope.HPVECs were divided into control group(NC group),exosomes group(NC+HPMSCs-EX group),injury group(LPS group)and exosome treatment group(LPS+HPMSCs-EX group).The effects of HPMSCs-EX on angiogenesis and migration of HPVECs were detected by angiogenesis experiment,migration experiment,scratch experiment,cell immunofluorescence experiment and Western blot experiment.Results Microscopically,HPMSCs were shaped like willow leaves,grew in a vortex,and had the ability to differentiate into fat,bone and cartilage.Transmission electron microscope showed that HPMSCs-EX were round cake with a diameter of less than 150 nm.Western blot showed that the expression of specific antigens CD9 and CD63 were positive,Calnexin was negative.HPMSCs-EX can be taken up by HPVECs.There was no significant difference in experimental results between NC group and NC+HPMSCs-EX group(P all>0.05).In LPS group,cell morphology was changed,intercellular connection was loose,angiogenesis and migration ability were decreased(P all<0.05),and VEGFA and Vimentin expression were down-regulated(P all<0.05).In LPS+HPMSCs-EX group,the cells were generally enlarged,some of them became round,bright and floating,but most of them remained in shape,their angiogenesis and migration ability were improved(P all<0.05),and the expressions of VEGFA and Vimentin were up-regulated(P all<0.05).Conclusion HPMSCs-EX may improve function of LPS induced HPVECs by promoting angiogenesis and migration.

关 键 词：人胎盘间充质干细胞 外泌体 肺血管内皮细胞 血管形成 迁移 

分 类 号：R563[医药卫生—呼吸系统]