益气活血复方治疗硬皮病肺部炎症与纤维化的作用机制:基于生物信息学与网络药理学方法  

The mechanisms by which Yiqi Huoxue compound treats pulmonary inflammation and fibrosis in scleroderma:Based on bioinformatics and network pharmacology

作　　者：胡梦华 谭乔瑞 濮伟霖 杜敏 屠文震 孙大燕 杨婧漪 顾逸飞 王久存[1] 孔祥贞 HU Menghua;TAN Qiaorui;PU Weilin;DU Min;TU Wenzhen;SUN Dayan;YANG Jingyi;GU Yifei;WANG Jiucun;KONG Xiangzhen(School of Life Sciences,Fudan University,Shanghai 200438,China;Guangdong-Hong Kong-Macao Greater Bay Area Institute of Precision Medicine,Guangzhou 511462,China;East China Hospital Affiliated to Fudan University,Shanghai 200040,China;Shanghai Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200438,China;Beijing Children′s Hospital,Capital Medical University,Beijing 100045,China;Second Affiliated Hospital of Soochow University,Suzhou 215004,China;Institute of Human Phenomics,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院,上海200438 [2]粤港澳大湾区精准医学研究院,广东广州511462 [3]复旦大学附属华东医院,上海200040 [4]上海中医药大学附属上海市中西医结合医院,上海200438 [5]首都医科大学附属北京儿童医院,北京100045 [6]苏州大学附属第二医院,江苏苏州215004 [7]复旦大学人类表型组研究院,上海200438

出　　处：《皮肤性病诊疗学杂志》2024年第5期303-312,共10页Journal of Diagnosis and Therapy on Dermato-venereology

基　　金：中国医学科学院院外创新单元项目(2019-I2M-5-066)。

摘　　要：目的通过生物信息学与网络药理学方法鉴定益气活血复方(YQHX)治疗硬皮病肺部炎症与纤维化的有效成分和关键分子,为硬皮病药物靶点开发提供依据。方法利用一次性气管灌注方法,使用博莱霉素诱导建立肺纤维化小鼠模型。将24只小鼠随机分成PBS对照组、博来霉素组、YQHX对照组、YQHX治疗组,每组6只,分别予以PBS或益气活血复方灌胃处理。利用HE染色、Masson染色方法验证YQHX对小鼠肺组织炎症及纤维化的治疗效果。从TCMSP数据库获取YQHX抗纤维成分丹参的活性成分及靶基因。整合PPI网络、GO/KEGG富集分析、分子对接等多种生物信息学分析,确定丹参治疗硬皮病的主要活性成分及关键靶点,分析相关信号通路。利用免疫组化分析小鼠肺组织标本中CD3抗体、p-PI3K抗体及p-AKT抗体表达情况,验证YQHX对炎症治疗及关键通路的影响。结果HE染色及Masson染色结果显示小鼠肺组织纤维化程度及炎症浸润下降,提示YQHX能够显著改善肺部纤维化、炎症及组织破坏程度。生物信息分析提示丹参中关键成分主要为木犀草素。PPI网络分析发现AKT1、EGFR、ESR1、TNF、IL6为丹参治疗硬皮病的核心靶点。KEGG富集发现基因主要富集在PI3K-AKT通路。分子对接分析发现木犀草素与4个关键靶点间存在潜在结合位点。免疫组化结果表明,YQHX显著降低博来霉素诱导的小鼠肺组织CD3+T细胞浸润、p-PI3K和p-AKT水平。结论YQHX对肺部炎症与纤维化具有显著的疗效。木犀草素可能是YQHX中关键有效分子,其可能通过靶向PI3K-AKT通路治疗硬皮病肺部炎症与纤维化。Objective To identify the effective components and key molecules of Yiqi Huoxu compound(YQHX)in the treatment of pulmonary inflammation and fibrosis in scleroderma,providing basis for the development of drug targets for scleroderma.Methods A mouse model of pulmonary fibrosis was established by a single tracheal perfusion of bleomycin.Twenty-four mice were randomly divided into four groups,i.e.intragastric PBS,bleomycin alone,intragastric Yiqi Huoxue compound,and control.HE staining and Masson staining were used to verify the therapeutic effect of Yiqi Huoxue compound on lung inflammation and fibrosis.The active components and target genes of anti-fiber salvia miltiorrhiza were obtained from TCMSP database.PPI network,GO/KEGG enrichment analysis,molecular docking and other bioinformatics analyses were integrated to identify the main active ingredients and key targets of salvia miltiorrhiza in the treatment of scleroderma,and to analyze the relevant signaling pathways.The expression levels of CD3 antibody,p-PI3K antibody and p-AKT antibody were analyzed by immunohistochemistry to verify the effects of Yiqi Huoxu compound on inflammation and key pathways.Results The results of HE staining and Masson staining showed that the severity of fibrosis and inflammatory infiltration were decreased,suggesting that Yiqi Huoxue compound could significantly improve the pulmonary fibrosis,inflammation and tissue damages.The key component of salvia miltiorrhiza was mainly luteolin.PPI network analysis showed that AKT1,EGFR,ESR1,TNF and IL6 were the core targets of salvia miltiorrhiza in the treatment of scleroderma.KEGG enrichment analysis showed that the genes were mainly enriched in the PI3K-AKT pathway.Molecular docking analysis showed the potential binding sites between luteolin and four key targets.The immunohistochemical results showed that YQHX significantly decreased bleomycin-induced CD3+T cell infiltration,p-PI3K and p-AKT levels in mouse lung tissue.Conclusions YQHX has significant therapeutic effects on lung inflammati

关 键 词：系统性硬化症 益气活血复方 丹参 肺纤维化 肺部炎症 

分 类 号：R28[医药卫生—中药学]