上皮细胞钠通道在博来霉素诱导小鼠肺纤维化中的作用机制  

Action mechanism of epithelial sodium channel in bleomycin-induced pulmonary fibrosis of mice

作　　者：丁炎 王琳[1] 刘妍宏 候亚鹏 聂宏光[1] DING Yan;WANG Lin;LIU Yanhong;HOU Yapeng;NIE Hongguang(Research Section of Stem Cells and Regenerative Medicine,College of Basic Medical Science,China Medical University,Shenyang,Liaoning 110122,China)

机构地区：[1]中国医科大学基础医学院干细胞与再生医学研究室,沈阳110122

出　　处：《重庆医学》2020年第21期3505-3509,共5页Chongqing medicine

基　　金：国家自然科学基金项目(81670010);辽宁省高等学校基本科研项目(LQNK201745)。

摘　　要：目的探讨上皮细胞钠通道(ENaC)在博来霉素诱导小鼠肺纤维化中的作用及相关机制,以及骨髓间充质干细胞条件培养基(BMSCs-CM)对ENaC的作用。方法将20只C57小鼠随机分为对照组和实验组。Masson染色观察小鼠肺纤维化程度;实时荧光定量-PCR(RT-PCR)和Western blot方法检测α-平滑肌肌动蛋白(α-SMA)、α/β/γ-ENaC mRNA的表达水平。将体外常规培养的肺泡Ⅱ型上皮细胞(AECⅡ)随机分为DMEM/F12组(CON组)、转化生长因子β1(TGF-β1)处理组(TGF组)、TGF-β1+BMSCs-CM组(T+CM组),培养24 h,采用Western blot方法检测α/γ-ENaC蛋白的变化。结果与对照组比较,实验组小鼠体重明显下降(P<0.05),死亡率为36.36%。实验组小鼠肺组织Masson染色观察到明显的肺细胞损伤和组织纤维化。RT-PCR和Western blot结果显示实验组小鼠TGF-β1、α-SMA mRNA和蛋白表达水平明显上升(P<0.05),α/β/γ-ENaC 3个亚基mRNA和蛋白的表达水平明显下降(P<0.05)。与CON组比较,TGF组小鼠的AECⅡ中α/γ-ENaC蛋白表达水平明显降低(P<0.05);与TGF组比较,T+CM组小鼠的AECⅡ中α/γ-ENaC蛋白表达水平明显升高(P<0.05)。结论BMSCs-CM对肺纤维化的保护作用可能与促进AECⅡ中ENaC的表达有关。Objective To investigate the role and related mechanism of epithelial sodium channel(ENaC)in bleomycin-induced pulmonary fibrosis of mice and the effects of bone marrow mesenchymal stem cells conditioned medium(BMSCs-CM)on ENaC.Methods Twenty C57 mice were randomly divided into the control group and experimental group.The pulmonary fibrosis degree was observed by Masson staining.The expression levels ofα-smooth muscle actin(α-SMA)andα/β/γ-ENaC were detected by using the real-time PCR(RT-PCR)and Western blot,respectively.The in vitro conventionally cultured typeⅡalveolar epithelial cells(AECⅡ)were randomly divided into the DMEM/F12 group(CON group),transforming growth factor-β1(TGF-β1)treatment group(TGF group)and TGF-β1+BMSCs-CM treatment group(T+CM group).The AECⅡwere cultured for 24 h.The Western blot method was used to detect the protein levels ofα/γ-ENaC.Results The body weight of the mice in the experimental group was significantly decreased compared with that in the control group,and the mortality rate was 36.36%.The obvious pulmonary cell injury and tissue fibrosis in the experimental group were observed by the Masson staining.The real-time PCR and Western blot results showed that the expression levels of TGF-β1 andα-SMA in the experimental group were increased(P<0.05),whereas the expression levels of 3 subunits ofα/β/γ-ENaC were decreased(P<0.05).Compared with CON group,the expression levels ofα/γ-ENaC protein in AECⅡof TGF group mice was decreased(P<0.05);compared with the simple TGF group,the expression levels ofα/γ-ENaC protein in the T+CM group were increased(P<0.05).Conclusion The protective effect of BMSCs-CM on pulmonary fibrosis may be related with promotion of ENaC expression in AECⅡ.

关 键 词：肺纤维化 博来霉素 上皮细胞钠通道 骨髓间充质干细胞条件培养基 上皮-间质转化 

分 类 号：R363.1[医药卫生—病理学]