TGF-β3拮抗TGF-β1诱导的FMT改善小鼠肺纤维化  

TGF-β3 Antagonizes TGF-β1-induced FMT to Improve Pulmonary Fibrosis in Mice

作　　者：覃林莽 林浩博 王婕颖 张光峰 徐婷 张晓 QIN Linmang;LIN Haobo;WANG Jieying(School of Medicine,South China University of Technology,Guangdong 510006,China)

机构地区：[1]华南理工大学医学院,广州510006 [2]南方医科大学附属广东省人民医院(广东省医学科学院)风湿免疫科,广州510080

出　　处：《医学研究杂志》2023年第4期31-35,72,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81771734);广东省基础与应用基础研究基金资助项目(2019A1515010047);广东省广州市科技计划项目(201804010343)。

摘　　要：目的 研究转化生长因子(transforming growth factor, TGF)-β3是否能够抑制TGF-β1诱导的人肺成纤维细胞(human lung fibroblast, HLF)间质转化。方法 在体外培养原代HLF进行细胞实验,分为3组:对照组:给予等体积超纯水;TGF-β1组:细胞给予TGF-β1(4ng/ml)处理HLF 24h;TGF-β3组:细胞给予20ng/ml的TGF-β3预处理24h后加入TGF-β1(4ng/ml)处理24h。Transwell实验检测细胞迁移和侵袭能力,Western blot法检测α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)和纤连蛋白(Fibronectin)蛋白的表达水平。在体内构建小鼠肺纤维化模型,分为3组:对照组:给予等体积0.9%氯化钠溶液气管滴注;模型组:博来霉素溶于0.9%氯化钠溶液,按2.0U/kg的浓度给予小鼠气管内滴注;TGF-β3干预组:于造模前1天,将TGF-β3(100μg/kg)给予小鼠尾静脉注射。HE和Masson染色法观察肺组织病理学变化,免疫荧光法检测α-SMA和Fibronectin蛋白的变化。结果 在体外,与TGF-β1组比较,TGF-β3组细胞迁移和侵袭能力降低,α-SMA和Fibronectin蛋白表达下调。在体内,与模型组比较,TGF-β3组小鼠肺组织病理变化改善,胶原沉积减少,α-SMA和Fibronectin蛋白荧光强度降低。结论 TGF-β3抑制TGF-β1引起的HLF间质转化,并改善博来霉素诱导的小鼠肺纤维化。Objective To investigate whether transforming growth factor(TGF)-β3 can inhibit TGF-β1-induced human lung fibroblasts(HLF)mesenchymal transformation.Methods In vitro,primary HLF were cultured for cell experiments and divided into 3 groups:control group:given equal volume of ultrapure water;TGF-β1 group:cells were given TGF-β1(4ng/ml)to treat HLF for 24h;TGF-β3 group:cells were pretreated with 20ng/ml TGF-β3 for 24h,and then treated with TGF-β1(4ng/ml)for 24h.Transwell assay was used to detect cell migration and invasion ability,and Western blot was used to detect the expression levels ofα-smooth muscle actin(α-SMA)and Fibronectin proteins.In vivo,the pulmonary fibrosis model of mice was constructed and divided into three groups:control group:received an equal volume of 0.9%sodium chloride solution by intratracheal instillation;model group:bleomycin was dissolved in 0.9%sodium chloride solution and given intratracheal instillation at a concentration of 2.0U/kg;TGF-β3 intervention group:1day before modeling,TGF-β3(100μg/kg)was injected into the tail vein of mice.The pathological changes in lung tissue were observed by HE and Masson staining,and the changes ofα-SMA and Fibronectin proteins were detected by immunofluorescence.Results In vitro,compared with the TGF-β1 group,the cell migration and invasion abilities of the TGF-β3 group were decreased,and theα-SMA and Fibronectin protein expression was down-regulated.In vivo,compared with the model group,the TGF-β3 group showed improved pathological changes in lung tissue,decreased collagen deposition,and decreased the fluorescence intensity ofα-SMA and Fibronectin protein.Conclusion TGF-β3 inhibits TGF-β1-induced mesenchymal transformation of HLF and improves bleomycin-induced pulmonary fibrosis in mice.

关 键 词：转化生长因子-Β3 转化生长因子-β1 间质转化 肺纤维化 

分 类 号：R563[医药卫生—呼吸系统]