TGF-β/Smad信号通路在类风湿性关节炎相关的间质性肺纤维化小鼠中的作用  

Role of TGF-β/Smad signaling pathway in pulmonary fibrosis after rheumatoid arthritis in mice

作　　者：付蓓蓓 陆雨琳 陈萌檬 鲍兰馨 刘楠楠 叶静[3] 陈晓宇[1] FU Bei-bei;LU Yu-lin;CHEN Meng-meng;BAO Lan-xin;LIU Nan-nan;YE Jin;CHEN Xiao-yu(School of Basic Medicine,Anhui Medical University,Hefei 230032,China;Dept of Clinical Medicine,Anhui Medical University,Hefei 230032,China;Dept of Respiratory Medicine,the Second Hospital of Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学基础医学院,安徽合肥230032 [2]安徽医科大学临床医学系,安徽合肥230032 [3]安徽医科大学第二附属医院呼吸内科,安徽合肥230601

出　　处：《中国药理学通报》2023年第3期483-488,共6页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金面上项目(No 2108085MH264);国家级大学生创新创业训练计划项目(No 202110366009);基础医临床合作提升项目(No 2019xkjT027)。

摘　　要：目的研究TGF-β/Smad信号通路在类风湿性关节炎(rheumatoid arthritis,RA)相关的间质性后肺纤维化小鼠中的作用。方法通过小鼠尾根部皮下给药完全弗氏佐剂(complete Freund′s adjuvant,CFA)与鸡Ⅱ型胶原蛋白(chickenⅡcollagen,Col-Ⅱ),构建RA小鼠模型。空白组给等量蒸馏水,对照组给等量冰醋酸(溶媒)。监测小鼠足趾肿胀程度(关节肿胀度与关节炎指数)评价小鼠建模情况;通过HE、Masson染色观察小鼠肺组织病理变化;免疫组化法观察肺间质细胞的胞质中TGF-β表达;采用化学发光法测量肺组织中羟脯氨酸水平;Western blot检测肺组织中Smad2、Smad3及p-Smad2、p-Smad3蛋白表达。结果与空白组、溶媒组相比,模型组小鼠关节肿胀、关节炎指数明显升高;给药21 d后,HE染色显示模型组肺间质有炎性改变,Masson染色显示模型组小鼠肺间质胶原纤维沉积,免疫组化染色显示模型组小鼠肺间质细胞的胞质中TGF-β表达升高,呈棕黄色;同时,与空白组、溶媒组相比,模型组肺组织匀浆显示羟脯氨酸表达明显升高;且进一步Western blot研究发现,与空白组、溶媒组相比,p-Smad2与p-Smad3在模型组中表达明显升高(P<0.05,P<0.01)。结论RA可导致肺组织纤维化,p-Smad2与p-Smad3表达上调,在肺纤维化和RA相关的间质性后肺纤维化中起重要作用。Aim To investigate the role of TGF-β/Smad signaling pathway in rheumatoid arthritis(RA)-associated postinterstitial pulmonary fibrosis in mice.Methods The mouse model of RA was constructed by subcutaneous administration of complete Freund′s adjuvant(CFA)and chicken II collagen(Col-Ⅱ)to the tail root of mice.The blank group was given the same amount of distilled water,and the control group was given the same amount of glacial acetic acid(solvent).The degree of toe swelling(joint swelling degree and arthritis index)was monitored to evaluate the mouse modeling.The pathological changes of mouse lung tissues were observed by HE and Masson staining.The expression of TGF-βin lung tissues were observed by immunohistochemical staining.The level of hydroxyproline in lung tissues was measured by chemiluminescence method.The expressions of Smad2,Smad3 and phosphorylated p-Smad2 and phosphorylated p-Smad3 in lung tissues were detected by Western blot.Results Compared with blank group and solvent group,the joint swelling and arthritis index of model group significantly increased.Twenty-one days after administration,HE staining showed inflammatory changes in lung interstitium of the model group,Masson staining showed collagen fiber deposition and obvious fibrosis in lung interstitium of the model group,and immunohistochemical staining showed that the expression of TGF-βin cytoplasm of lung interstitial cells of the model group increased,which was brown and yellow.Meanwhile,hydroxyproline was significantly raised in lung tissue homogenate of the model group.Further WB analysis showed that compared with blank group and solvent group,the expression of p-Smad2 and p-Smad3 in lung tissues of the model group was significantly up-regulated(P<0.05,P<0.01).Conclusions RA can give rise to pulmonary fibrosis,and the expressions of p-Smad2 and p-Smad3 are up-regulated,which is be pivotal in pulmonary fibrosis and RA-related post-interstitial pulmonary fibrosis.

关 键 词：类风湿性关节炎 间质性肺病 肺纤维化 TGF-Β/SMAD信号通路 p-Smad2 p-Smad3 

分 类 号：R332[医药卫生—人体生理学] R322.35[医药卫生—基础医学] R341R563.13R593.22R977.6