Network pharmacology-based research of mechanism of stemona and Fritiliariae cirrhosae bulbus in treatment of pulmonary tuberculosis
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 H37Ra 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.
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