人脂肪干细胞外泌体对烟熏诱导小鼠炎症和肺部损伤的治疗作用  

Therapeutic effect and underlying mechanism of exosomes from human adipose-derived stem cells on smoke-induced inflammation and lung injury in mice

作　　者：连细华 苏小珊 叶祥嘉 王大春 曾奕明[2,3] 朱志兴[2,3] LIAN Xihua;SU Xiaoshan;YE Xiangjia;WANG Dachun;ZENG Yiming;ZHU Zhixing(Department of Medical Imaging,Second Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian Province,362000,China;Department of Pulmonary and Critical Care Medicine,Respiratory Medicine Centre of Fujian Province,Second Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian Province,362000,China;Stem cell Research Lab,Second Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian Province,362000,China)

机构地区：[1]福建医科大学附属第二医院医学影像科,福建泉州362000 [2]福建医科大学附属第二医院呼吸与危重症医学科,福建省呼吸医学中心,福建泉州362000 [3]福建医科大学附属第二医院干细胞实验室,福建泉州362000

出　　处：《第三军医大学学报》2021年第20期2180-2188,共9页Journal of Third Military Medical University

基　　金：国家重点研发计划(2016YFC1304);国家自然科学基金面上项目(82070021)。

摘　　要：目的探讨人脂肪干细胞(human adipose-derived stem cells,hADSCs)外泌体对烟熏诱导慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)小鼠炎症反应和肺部损伤的治疗作用和机制,每组8只。方法从hADSCs的培养上清液中分离外泌体。C57bl/6小鼠按照随机数字表法分为对照组、COPD组和外泌体组,每组8只。烟熏4周构建COPD模型,外泌体组小鼠经气道滴注30μL外泌体(蛋白浓度100 mg/L),其余2组小鼠仅滴注等体积PBS,4周后处死小鼠并收集标本。ELISA法检测血清TNF-α、IL-6和趋化因子CXCL1水平;免疫荧光测定肺组织焦亡相关蛋白Caspase-1、NLRP3和ACS水平;Western blot测定肺组织凋亡相关蛋白Bcl-2、Bax、Caspase-3和Caspase-9的水平;HE和PAS染色观察肺组织病变和粘液分泌情况。结果与对照组比较,COPD组小鼠体质量增长缓慢;血清TNF-α、IL-6和CXCL1均升高;肺组织Caspase-1、NLRP3和ACS表达增多,Bcl-2表达减少,而Bax、Caspase-3和Caspase-9表达均增加(P<0.05)。与COPD组比较,外泌体组小鼠体质量增长较快;血清TNF-α、IL-6和CXCL1均降低(P<0.05);肺组织Caspase-1、NLRP3和ACS表达减少,Bcl-2表达增加,而Bax、Caspase-3和Caspase-9表达减少(P<0.05)。COPD组小鼠肺组织破坏明显,可见大量粘液分泌;外泌体治疗后小鼠肺组织损伤减轻,粘液分泌明显减少。结论hADSCs外泌体可减轻烟熏诱导的炎症水平,抑制肺组织焦亡和凋亡发生,减少气道粘液分泌,促进肺损伤修复从而治疗COPD。Objective To investigate the therapeutic effects of exosomes from human adipose-derived stem cells(hADSCs-Exos)on the inflammation and lung injury in mice with smoke-induced chronic obstructive pulmonary disease(COPD),and explore the underlying mechanism.Methods Exosomes were isolated from culture supernatant of hADSCs.A total of 24 male C57 bl/6 mice were randomly divided into 3 groups(n=8 for each group),namely the control,COPD and exosomes-treated group,respectively.The mice of the latter 2 groups were exposed to cigarette smoke for 4 weeks to establish the COPD model.The exosomes-treated group received intratracheal instillation of 30μL hADSCs-Exos(100 mg/L),while the control and COPD groups were treated with 30μL PBS.Their body weights were monitored weekly for 4 weeks before being sacrificed.The concentrations of TNF-α,IL-6 and chemokine CXCL1 were detected by ELISA.The levels of pyroptosis-related proteins in the lungs like Caspase-1,ASC and NLRP3 were determined using immunofluorescence assay,and the expression levels of apoptosis-related proteins Bcl-2,Bax,Caspase-3 and Caspase-9 were measured by Western blotting.In addition,pathological abnormalities of the lungs and mucus secretion in the airway were observed after HE and PAS staining.Results Compared with the control group,the mice in the COPD group had lower body weight and higher concentrations of TNF-α,IL-6 and CXCL1,up-regulated expression of Caspase-1,ASC and NLRP3 but down-regulated level of Bcl-2 in the lungs,and those of Bax,Caspase-3 and Caspase-9 were increased(all P<0.05).In comparison with the COPD group,the exosomes-treated group had higher body weight,lower levels of TNF-α,IL-6 and CXCL1,as well as down-regulated Caspase-1,ASC,NLRP2,Bax,Caspase-3 and Caspase-9,while the expression of Bcl-2 was increased(all P<0.05).Histologically,lung tissues were severely damaged and pulmonary mucus secretion was obvious in the COPD group,whereas,the injuries were relieved with diminished mucus production in the exosomes-treated group.Conclusion hA

关 键 词：脂肪干细胞外泌体 炎症 细胞焦亡 细胞凋亡 粘液分泌 肺损伤 

分 类 号：R-332[医药卫生] R329.2