摘要
目的运用网络药理学方法探讨百部联合川贝母干预肺结核病的作用机制,并通过细胞实验验证其核心靶点的作用。方法首先通过关联规则法筛选出中药对百部、川贝母。然后利用TCMSP数据平台,筛选百部和川贝母的主要化学成分,从GeneCards数据库中检索肺结核病对应的基因,通过String数据库构建蛋白质互作用网络,发现关键靶基因,利用Cytoscape软件进行GO和KEGG生物富集分析。最后构建结核分枝杆菌感染小鼠单核巨噬细胞的细胞实验,利用FCM(flow cytometry)技术检测细胞凋亡、Western blot技术检测靶点PPARG的表达。结果筛选出与肺结核病相关靶点2320个,维恩映射得到27个交集基因,蛋白质互作用网络分析得到关键靶基因7个,靶点PPARG尚未得到验证,生物富集分析,得到76条GO term,20条pathway。构建结核分枝杆菌菌株H_(37)Ra感染小鼠单核巨噬细胞RAW264.7的细胞实验模型,验证靶基因PPARG的蛋白表达量差异。结论利用关联规则法能发现中药方剂中的有效中药组合,进一步通过网络药理学分析百部、川贝母联合治疗肺结核病的生物进程和信号通路,涉及炎症与免疫、增殖与凋亡、氧化应激、神经传导和神经抑制等多靶点作用。结核分枝杆菌菌株H_(37)Ra感染小鼠单核巨噬细胞RAW264.7的实验结果显示:基因PPARG抗体的表达上调。验证了百部、川贝母治疗肺结核的关键基因,在分子生物学层面为中药联合治疗肺结核提供依据。
Objective To explore the anti-pulmonary tuberculosis mechanism of stemona and Fritiliariae cirrhosae bulbus based on network pharmacology and verify the role of its key targets by cell experiments.Methods Using the association rule method,the drug combination stemona and Fritiliariae cirrhosae bulbus was screened.The main chemical constituents of stemona and Fritiliariae cirrhosae bulbus were identified based on the traditional Chinese medicine systems pharmacology database and analysis platform.According to the pharmacokinetic characteristics of traditional Chinese medicine,pulmonary tuberculosis related gene targets were screened in the Genecards database.Protein interaction network was generated based on the String database to discovery key target genes.GO and KEGG enrichment analyses were then performed.Cell experiments were performed to verify cell apoptosis by flow cytometry(FCM).The expression of PPARG protein was detected by Western blot.Results A total of 2320 gene targets related to pulmonary tuberculosis were screened,21 intersecting genes were obtained by Venn mapping,and 7 key target genes were identified through protein interaction network analysis.GO enrichment analysis yielded 76 GO terms,and KEGG pathway enrichment analysis yielded 20 pathways.A cell model was developed using Mycobacterium tuberculosis strain H_(37)Ra infected mouse monocyte-macrophage RAW264.7 cells to determine the rate of apoptosis by FCM and examine the protein expression of PPARG by WB analysis.Conclusion The association rule method could be used to find the effective combination of traditional Chinese medicine in traditional Chinese medicine formulae.The biological processes and signal pathways involved in the therapeutic effects of stemona and Fritiliariae cirrhosae bulbus on pulmonary tuberculosis habe been identified through network pharmacology,which involve multi-target effects such as inflammation and immunity,proliferation and apoptosis,oxidative stress,and nerve conduction and suppression.Cell experiments suggested that PPARG is upregulated and may be the key target of stemona and Fritiliariae cirrhosae bulbus in the treatment of pulmonary tuberculosis.These results providing a new avenue for further exploring the molecular mechanism of traditional Chinese medicine combinations.
作者
郭文秀
吴胜男
王小芸
张敏
Guo Wenxiu;Wu Shengnan;Wang Xiaoyun;Zhang Min(School of Management,Shanxi Medical University,Taiyuan 030001,China;Shanxi Academy of Traditional Chinese Medicine,Taiyuan 030001,China;School of pharmacy Shanxi Medical University,Taiyuan 030001,China)
出处
《中华临床医师杂志（电子版）》
CAS
北大核心
2023年第3期335-342,共8页
Chinese Journal of Clinicians(Electronic Edition)
基金
国家自然科学基金项目(71804102)
关键词
百部
川贝母
肺结核
网络药理学
Stemona
Fritiliariae cirrhosae bulbus
Pulmonary tuberculosis
Network pharmacology
