关键词: 隐孢子虫病, 苦参汤, 网络药理学, 分子对接, 协同作用机制

Abstract: Objective　To explore the potential targets and synergistic mechanisms of Kushen Decoction for the treatment of cryptosporidiosis using network pharmacology and molecular docking methods. Methods　The main active ingredients of Kushen Decoction were captured from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the Universal Protein Resource (UniProt) database, and the potential targets were predicted. In addition, the active ingredients of Kushen Decoction that were not included in the TCMSP database were retrieved in CNKI, WanFang Data, CBM, PubMed and Web of Science databases, and the target genes of all supplemented active ingredients were predicted using the online TargetNet database. Network construction and analysis were performed using the Cytoscape software, and cryptosporidiosis⁃related targets were retrieved in the Comparative Toxicogenomics Database and GeneCards database. The protein⁃protein interaction (PPI) network was created using the STRING database, and the DAVID database was used for GO enrichment and KEGG pathway analyses. The tissue distribution of key targets was investigated using the BioGPS database, and the AutoDockTools software was employed to verify the molecular docking results. Results　A total of 38 active ingredients of Kushen Decoction were screened, and the core ingredients included quercetin, (+)⁃14α⁃hydroxymatrine and apigenin. A total of 831 targets of Kushen Decoction and 512 cryptosporidiosis⁃related targets were predicted, and PPI network analysis revealed 69 key targets, including AKT1, TNF and IL⁃6. There were 303 biological processes, 46 molecular functions and 29 cellular components involved in the treatment of cryptosporidiosis with Kushen Decoction, and 13 KEGG pathways played a therapeutic role in the synergistic mechanisms of multiple targets, such as Toll⁃like receptor (TLR), nuclear factor kappa B(NF)⁃ĸB, nucleotide binding oligomerization domain like receptor (NLR) signal pathways. The core targets were mainly distributed in the hematologic and immune systems. Molecular docking analysis showed that the binding energy between active ingredients and key targets were all less than 0 kJ/mol, indicating the strong binding of ligands to receptors. Conclusions　The active ingredients of Kushen Decoction, such as quercetin, (+)⁃14α⁃hydroxymatrine and apigenin, may act on targets like AKT1, TNF, IL⁃6 to modulate TLR, NLR and NF⁃κB signaling pathways to play a synergistic role in the treatment of cryptosporidiosis in the hematologic and immune system.

Key words: Cryptosporidiosis, Kushen Decoction, Network pharmacology, Molecular docking, Synergistic mechanism