Toll样受体4在骨髓间充质干细胞外泌体调控上皮间质转化抑制急性呼吸窘迫综合征后肺纤维化中的作用机制  

The mechanism of TLR4 in the regulation of EMT by extracellular vesicles of bone marrow mesenchymal stem cells in inhibiting pulmonary fibrosis after ARDS

作　　者：刘树安 陈天恩 经慧 武焱旻[2] 薛欢家 吴滢 王凯[3] LIU Shuan;CHEN Tianen;JING Hui;WU Yanmin;XUE Huanjia;WU Ying;WANG Kai(Pulmonary and Critical Care Medicine,Xuzhou Cancer Hospital,Xuzhou,Jiangsu221009,China;Pulmonary and Critical Care Medicine,Xuzhou Central Hospital,Xuzhou Clinical School of Xuzhou Medical University,Xuzhou,Jiangsu221009,China;Department of Anesthesiology,Xuzhou Central Hospital,Xuzhou Clinical School of Xuzhou Medical University,Xuzhou,Jiangsu221009,China)

机构地区：[1]徐州市肿瘤医院呼吸与危重症医学科,江苏徐州221009 [2]徐州市中心医院/徐州医科大学徐州临床学院呼吸与危重症医学科,江苏徐州221009 [3]徐州市中心医院/徐州医科大学徐州临床学院麻醉科,江苏徐州221009

出　　处：《临床肺科杂志》2024年第11期1633-1637,共5页Journal of Clinical Pulmonary Medicine

基　　金：江苏省自然科学基金面上项目(No.BK20171172);江苏省徐州市科技计划面上项目(No.KC21055);江苏省徐州市医学重点人才项目(No.XWRCHT20220051)。

摘　　要：目的探究Toll样受体4(TLR4)在骨髓间充质干细胞(BMSCs)来源的外泌体(Exo)调控上皮间质转化(EMT)抑制急性呼吸窘迫综合征(ARDS)后肺纤维化中的作用机制。方法提取、培养Ⅱ型肺泡上皮细胞,构建ARDS后肺纤维化模型,进行TLR4慢病毒转染和基因干扰,分为control组、NC TLR4^(+)组、TLR4^(+)组、NC TLR4^(-)组、TLR4^(-)组,转染完成后进行实时荧光定量(Q-PCR)验证转染效果,明确转染效果后,进行后续实验,按是否加入Exo干预分为模型组、Exo治疗组、空载体+Exo治疗组、高表达TLR4^(+)Exo治疗组、对照序列+Exo治疗组、低表达TLR4^(+)Exo治疗组,48小时后收集细胞,Western Blot检测TLR4效应蛋白MyD88、NF-κB、TGF-β1及EMT相关蛋白E-cadherin、α-SMA表达情况。结果比较各组TGF-β1、MyD88、NF-κB表达发现,Exo治疗组低于模型组;低表达TLR4^(+)Exo治疗组低于Exo治疗组;高表达TLR4^(+)Exo治疗组高于Exo治疗组(P<0.05)。其他组间比较差异无统计学意义,(P>0.05)。结论BMSCs-Exo可以通过调控TLR4/MyD88/NF-κB信号通路及TGF-β1的表达调控EMT抑制ARDS后肺纤维化。Objective To investigate the mechanism of Toll-like receptor 4(TLR4)in the regulation of epithelial mesenchymal transition(EMT)by exosomes derived from bone marrow mesenchymal stem cells(BMSCs)in inhibiting pulmonary fibrosis after acute respiratory distress syndrome(ARDS).Methods Type II alveolar epithelial cells were extracted and cultured to construct the pulmonary fibrosis model after ARDS.TLR4 lentiviral transfection and gene interference were performed,and the cells were divided into a control group,NC TLR4^(+)group,TLR4^(+)group,NC TLR4^(-)group,TLR4^(-)group,Real-time fluorescence quantification(Q-PCR)was performed after transfection to verify the transfection effect.After the transfection effect was determined,follow-up experiments were conducted,which were divided into model group,Exo treatment group,empty carrier+Exo treatment group,high-expression TLR4^(+)Exo treatment group,control sequence+Exo treatment group,low-expression TLR4^(+)Exo treatment group according to whether Exo intervention was added.Cells were collected 48 hours later.Western Blot analysis was performed to detect the expression of TLR4 affecting proteins MyD88,NF-κB,TGF-β1,and EMT-related proteins E-cadherin andα-SMA.Results The degree of the expression of TGF-β1,MyD88,and NF-κB among all groups were compared,and the Exo treatment group was lower than the model group.The low expression TLR4^(+)Exo group was lower than that of the Exo group.The high expression of TLR4^(+)in the Exo group was higher than that in the Exo group(P<0.05).There was no significant difference between the other groups.Conclusion BMSCs-Exo regulate EMT to inhibit pulmonary fibrosis after ARDS by regulating TLR4/MyD88/NF-κB signaling pathway and the expression of TGF-β1.

关 键 词：肺纤维化 上皮间质转化 外泌体 

分 类 号：R563.8[医药卫生—呼吸系统]