间充质干细胞来源外泌体对HDM刺激的气道上皮细胞炎症及EMT的影响  

Effects of mesenchymal stem cell-derived exosomes on inflammation and EMT of airway epithelial cells stimulated by HDM

作　　者：胡倩[1] 李国荣[1] 侯晨辉[1] 王建英[1] 寇应琳[1] HU Qian;LI Guorong;HOU Chenhui;WANG Jianying;KOU Yinglin(Zhengzhou Railway Vocational and Technical College,Zhengzhou 450000,China)

机构地区：[1]郑州铁路职业技术学院,郑州450000

出　　处：《中国免疫学杂志》2023年第3期560-565,共6页Chinese Journal of Immunology

摘　　要：目的:探究间充质干细胞来源外泌体(MSC-EXO)对屋尘螨(HDM)刺激的气道上皮细胞炎症及上皮-间质转化(EMT)的影响。方法:从人脐带中分离MSCs(hUCMSCs),经过分选和鉴定后分离提取MSC-EXO。透射电镜和Western blot检测外泌体形态及标志蛋白CD63、CD81、TSG101表达。将MSC-EXO与HDM刺激的16HBE细胞共培养后,观察细胞形态变化;MTT检测细胞增殖能力;流式细胞术检测细胞凋亡率;qRT-PCR检测细胞炎症因子TNF-α和IL-6 mRNA水平;荧光探针法检测细胞活性氧(ROS)水平;Western blot检测EMT相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(VIM)、α平滑肌肌动蛋白(α-SMA)表达。结果:hUCMSC中CD73、CD90和CD105表达呈阳性,CD34、CD45和HLA-DR表达呈阴性;电子透射显微镜下MSC-EXO呈囊泡状,直径30~200 nm,表达CD63、CD81、TSG101。与对照组相比,HDM组16HBE细胞变长、变尖呈长梭形,细胞凋亡率、ROS、TNF-α和IL-6 mRNA水平、N-cadherin、VIM、α-SMA表达显著升高,E-cadherin表达显著降低(P<0.05);与HDM组相比,50、100μg/ml EXO组细胞形态接近正常细胞,细胞凋亡率、ROS、TNF-α和IL-6 mRNA水平、N-cadherin、VIM、α-SMA表达明显降低,E-cadherin表达明显升高(P<0.05)。结论:MSC-EXO可减轻HDM诱导的气道上皮细胞炎症,抑制EMT过程。Objective:To investigate effect of mesenchymal stem cell-derived exosomes(MSC-EXO)on airway epithelial inflammation and epithelial-mesenchymal transition(EMT)induced by house dust mite(HDM).Methods:MSCs were isolated from human umbilical cord(hUCMSCs),MSC-EXO was isolated after sorting and identification.Morphology of exosomes and expressions of CD63,CD81 and TSG101 marker proteins were detected by transmission electron microscope and Western blot.MSC-EXO and HDM stimulated 16HBE cells were co-cultured to observe morphological changes;cell proliferation was detected by MTT;apoptosis rate was detected by flow cytometry;mRNA levels of cytokines TNF-αand IL-6 were detected by qRT-PCR;level of reactive oxygen species(ROS)was detected by fluorescence probe;expressions of EMT related proteins E-cadherin,N-cadherin,vimentin(VIM)andα-smooth muscle actin(α-SMA)were detected by Western blot.Results:CD73,CD90 and CD105 expressions in hUCMSC were positive,while expressions of CD34,CD45 and HLA-DR were negative;under electron microscope,MSC-EXO was vesicular,30~200 nm in diameter,and expressed CD63,CD81 and TSG101.Compared with control group,16HBE cells in HDM group became longer,sharper and longer fusiform,apoptosis rate,ROS,levels of TNF-αand IL-6 mRNA,and expressions of N-cadherin,VIM andα-SMA were significantly increased,expression of E-cadherin was significantly decreased(P<0.05);compared with HDM group,morphology of cells in 50μg/ml and 100μg/ml EXO groups were closer to normal cells,apoptosis rate,ROS,levels of TNF-αand IL-6 mRNA,and expressions of N-cadherin,VIM andα-SMA were significantly decreased,E-cadherin expression was significantly increased(P<0.05).Conclusion:MSC-EXO can reduce inflammation of airway epithelial cells induced by HDM and inhibit EMT process.

关 键 词：间充质干细胞来源外泌体 气道上皮细胞 哮喘 气道炎症 上皮-间质转化 

分 类 号：R392.11[医药卫生—免疫学]