基于网络药理学研究升陷汤合金水六君煎治疗矽肺的作用机制  

Research on the mechanism of shengxian and jinshuiliujun decoction in treating silicosis based on network pharmacology

作　　者：汤旖雯 张笑璇 吴冰冰 赵丽媛 沈曦 沈福海 Tang Yiwen;Zhang Xiaoxuan;Wu Bingbing;Zhao Liyuan;Shen Xi;Shen Fuhai(School of Public Health,North China University of Science and Technology,Hebei Province Key Laboratory of Occupational Health and Safety for Coal Industry,Key Laboratory of Organ Fibrosis in Hebei Province,Tangshan 063210,China)

机构地区：[1]华北理工大学公共卫生学院,河北省煤矿卫生与安全实验室,河北省器官纤维化重点实验室,唐山063210

出　　处：《中华劳动卫生职业病杂志》2024年第7期487-497,共11页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金(U21A20334)。

摘　　要：目的运用网络药理学方法探讨升陷汤合金水六君煎的活性成分,对其治疗矽肺机制进行实验验证。方法于2023年5月,通过中药系统药理学数据库与分析平台(TCMSP)获取升陷汤合金水六君煎中药物有效成分和靶点,通过人类基因的综合数据库Genecards、疾病相关基因与突变位点数据库(DisGeNET)、比较毒物基因组学数据库(CTD)等筛选矽肺疾病靶点,将筛选的药物靶点和疾病靶点取交集,获取升陷汤合金水六君煎治疗矽肺的靶点集。通过STRING数据库对靶点集进行蛋白相互作用(PPI)网络分析,并筛选核心靶基因。基于Metascape数据库对交集基因进行GO富集分析和KEGG通路分析,并对升陷汤合金水六君煎关键作用成分与靶点进行分子对接验证。将24只SPF级成年雄性SD大鼠随机分为对照组、模型组、中药干预组,每组8只。采用1 ml二氧化硅混悬液(50 mg/ml)非气管暴露一次性制备染尘大鼠模型,中药干预组于第2天给予升陷汤合金水六君煎灌胃[6 g/(kg·d)]。观察染尘后28 d各组大鼠肺部CT情况,取大鼠肺组织制备石蜡切片并进行苏木素-伊红(HE)及马松(Masson)染色,蛋白免疫印迹(Western blot)法验证升陷汤合金水六君煎干预后28 d大鼠肺组织核心靶点相关蛋白表达,用单因素方差分析比较组间蛋白的表达差异。结果共筛选出升陷汤合金水六君煎中205个有效活性成分,活性化合物3345个,对应靶点281个,其中与矽肺相关靶点240个。丝氨酸/苏氨酸激酶1(AKT1)、肿瘤蛋白p53(TP53)、肿瘤坏死因子(TNF)、白细胞介素(IL)6可能是升陷汤合金水六君煎治疗矽肺的关键靶点。富集分析并根据P值筛选出30条GO条目和20个潜在信号通路,主要包括核因子κB(NF-κB)、丝裂原活化蛋白激酶(MAPK)和癌症等信号通路。分子对接得出升陷汤合金水六君煎活性化合物与核心靶点蛋白结合性较好,其中结合能最强的是β-谷甾醇和TNF-α(-10.ObjectiveTo explore the active ingredients of shengxian and jinshuiliujun decoction with the method of network pharmacology,and to verify the experimental mechanism of its treatment of silicosis.MethodsIn May 2023,the active ingredients and targets of drugs in shengxian and jinshuiliujun decoction were obtained through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database.The target of silicosis disease was screened by databases such as Genecards,Disease Gene Network(DisGeNET),Comparative Toxicogenomics Database(CTD),etc.The screened drug targets and disease targets were intersected to obtain the target set of shengxian and jinshuiliujun decoction for the treatment of silicosis.Protein-protein interaction(PPI)network analysis was performed on the target set through STRING database,and core target genes were screened.GO enrichment analysis and KEGG pathway analysis of intersection genes were performed based on Metascape database,and molecular docking verification of key components and targets of shengxian and jinshuiliujun decoction was carried out.Twenty-four adult male SD rats with SPF grade were randomly divided into control group,model group and TCM intervention group,with 8 rats in each group.The dust-stained rat model was prepared by non-tracheal exposure of 1 ml silica suspension(50 mg/ml)in one go,and TCM intervention group was given shengxian and jinshuiliujun decoction[6 g/(kg·d)]on the second day.The CT of the lungs of each group was observed 28 days after the dust-stained rat model.Paraffin sections of rat lung tissues were prepared and stained with Hematoxylin-Eosin(HE)and Masson.Western blot was used to verify the expression of core target-related proteins in rat lung tissues after the intervention of shengxian and jinshuiliujun decoction for 28 days,and the differences in protein expression between groups were compared by one-way analysis of variance.ResultsA total of 205 active ingredients and 3345 active compounds were selected from shengxian and jins

关 键 词：矽肺 升陷汤 金水六君煎 网络药理学 分子对接 大鼠 肺纤维化 

分 类 号：R285[医药卫生—中药学]