穿山龙总皂苷通过TGF-β/Smad信号通路减轻类风湿关节炎相关间质性肺病的肺纤维化  

Total Saponins from Rhizoma Dioscoreae Nipponicae Attenuate Pulmonary Fibrosis in Rheumatoid Arthritis-associated Interstitial Lung Disease through TGF-β/Smad Signaling Pathway

作　　者：吴晓东 徐胜梅 杨智华 梅丽艳 王茂杰 陈秀敏 韩凌 黄闰月 WU Xiaodong;XU Shengmei;YANG Zhihua;MEI Liyan;WANG Maojie;CHEN Xiumin;HAN Ling;HUANG Runyue(The Second Affiliated Hospital,Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Chinese Medicine),Guangzhou 510120,Guangdong,China;Guangzhou University of Chinese Medicine,Guangzhou 510140,Guangdong,China;Guangdong Provincial Key Laboratory of Research on Emergency in TCM,Guangzhou510120,Guangdong,China;State Key Laboratory of Dampness Syndrome of Chinese Medicine,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong,China;Guangdong-Hong Kong-Macao Joint lab on Chinese Medicine and Immune Disease Research,Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong,China;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome,Guangzhou 510120,Guangdong,China)

机构地区：[1]广州中医药大学第二附属医院,广东广州510120 [2]广州中医药大学,广东广州510140 [3]广东省中医急症研究重点实验室,广东广州510120 [4]广州中医药大学第二附属医院省部共建中医湿证国家重点实验室,广东广州510120 [5]广州中医药大学粤港澳中医药与免疫疾病研究联合实验室,广东广州510120 [6]广东省中医证候临床研究重点实验室,广东广州510120

出　　处：《中华中医药学刊》2023年第1期99-102,I0033-I0035,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81774218);广东省自然科学基金项目(2021A1515011477,2021A1515011593);广东省科学技术厅项目(2016A020226041);广东省中医院临床研究项目(YN10101906,YN2021RD01);广州中医药大学第二附属医院中医湿证国家重点实验室项目(SZ2021ZZ35)。

摘　　要：目的探讨穿山龙总皂苷(total saponins from rhizoma dioscoreae nipponicae,TSRDN)在类风湿关节炎间质性肺病(rheumatoid arthritis-associated interstitial lung disease,RA-ILD)纤维化发病机制中的治疗可能性和潜在机制。方法在DBA/1小鼠胶原诱导关节炎(collagen-induced arthritis,CIA)模型中气管内滴注博来霉素(bleomycin,BLM)建立新的体内RA-ILD模型和转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人肺成纤维细胞(HFL1)体外模型中研究穿山龙总皂苷对RA-ILD肺纤维化的药理作用和可能机制。苏木精-伊红染色(HE)观察小鼠踝关节和肺组织的病理变化,Masson染色用于检查肺组织的纤维化程度,MTT法用于检测穿山龙总皂苷对细胞活力的影响,利用实时荧光定量PCR(RT-qPCR)、蛋白免疫印迹法(WB)、免疫组织化学法(IHC)和免疫荧光(IF)等方法分析TGF-β/Smad途径的基因和蛋白表达情况。结果穿山龙总皂苷可以改善BLM诱导的RA-ILD小鼠肺功能,减轻肺组织和关节的病理变化。在体外,穿山龙总皂苷抑制TGF-β1诱导的HFL1细胞中I型胶原蛋白(Collagen I)和α-平滑肌肌动蛋白(α-SMA)的蛋白和mRNA表达。此外,穿山龙总皂苷在体外和体内降低了由TGF-β1激活的p-Smad2/3的表达。结论穿山龙总皂苷可以减轻RA-ILD的肺纤维化,其机制可能是通过抑制TGF-β/Smad通路实现的。Objective To investigate the therapeutic possibility and the underlying mechanism of the total saponins from Rhizoma Dioscoreae Nipponicae(TSRDN)in fibrosis of rheumatoid arthritis-associated interstitial lung disease(RA-ILD).Methods In this article,we used a new in vivo RA-ILD model established by intratracheal instillation of bleomycin(BLM)in DBA/1 mice with collagen-induced arthritis(CIA)and an in vitro model induced by transforming growth factor-β1(TGF-β1)in human lung fibroblasts(HFL1)cells to investigate the pharmacological effects and possible mechanisms of the TSRDN against pulmonary fibrosis in RA-ILD.Hematoxylin and eosin(H&E)staining was performed to evaluate pathological changes in ankle joints and the lung tissue.Masson’s trichrome staining was used to examine the degree of fibrosis in lung tissue.MTT assay was used to detect the impact of TSRDN on cell viability.Real-time quantitative PCR(RT-qPCR),western blot,immunohistochemistry and immunofluorescence were performed to analyze the genes and proteins expression of TGF-β/Smad pathway.Results We observed that TSRDN could improve pulmonary function induced by BLM.Moreover,TSRDN relieved pathological changes of lung and joints.In vitro,we found that TSRDN inhibited TGF-β1-induced production of type I collagen protein andα-SMA in HFL1 cells.And TSRDN suppressed TGF-β1-induced expression of type I collagen(COL1A1)mRNA andα-SMA(ACTA2)mRNA.In addition,TSRDN reduced the expression of p-Smad2/3 activated by TGF-β1 in vitro and in vivo.Conclusions These findings illustrated TSRDN can reduce lung fibrosis of RA-ILD and its mechanism may be through inhibition of TGF-β/Smad path.

关 键 词：类风湿关节炎 间质性肺病 穿山龙总皂苷 TGF-Β1 SMAD2/3 

分 类 号：R285.5[医药卫生—中药学]