关键词: 芹菜素, 奶牛, 热应激, 低氧, 网络药理学, 分子对接

Abstract:

【Objective】 The study aimed to predict the mechanisms of apigenin in relieving heat stress and hypoxia stress in dairy cows by using network pharmacology and molecular docking so as to provide a reference for the application of apigenin.【Method】 Firstly, the targets related to apigenin, heat stress, and hypoxia stress were obtained from TCMSP and GeneCards databases, and apigenin targets in relieving dual stress were obtained by Venn intersection. The protein-protein interaction (PPI) network was constructed using the STRING database, and the core targets were identified by analyzing the network node centrality using the CytoNCA plugin in Cytoscape 3.9.1 software. Autodock vina program was used to perform molecular docking of apigenin with the identified targets. Perform gene ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the top 30 ranked targets based on degree centrality using Functional Annotation, the analysis tool of David database. Additionally, Molecular complex detection analysis (MCODE) of the above targets yielded the core gene clusters, then GO and KEGG analysis were performed. Apigenin-dual stress targets-signaling pathway network model were constructed through the KEGG pathway enrichment results and core target information. Finally, apigenin was docked to the core targets using the Autodock vina program, and the results were visualized using the Pymol software and the PLIP website.【Result】 The TCMSP database retrieved 68 targets of apigenin, 6 661 hypoxia-related genes and 9 046 heat stress-related genes were obtained in GeneCards database, and 56 apigenin targets in relieving dual stress were obtained. The number of nodes in the target PPI network was 56, the number of edges was 436, the average node degree value was 15.6, the mean local clustering coefficient was 0.728, and PPI enrichment P value was less than 1.0×10^-16. The core targets were AKT1 (RAC-alpha serine/ threonine-protein kinase), TP53 (Cellular tumor antigen p53), TNF (Tumor necrosis factor), CASP3 (Caspase-3), INS (Insulin), BCL2L1 (Bcl-2-like protein 1), VEGFA (Vascular endothelial growth factor A), HIF1A (Hypoxia-inducible factor 1-alpha), PTGS2 (Prostaglandin G/H synthase 2), and SREPINE1 (Plasminogen activator inhibitor 1). The molecular docking results showed that apigenin could be stably bound to the above core targets. A total of 54 Biological Processes (BP), 6 Molecular Functions (MF), and 11 Cellular Components (CC) were obtained by GO functional enrichment analysis (P<0.01), and KEGG were enriched to 98 signaling pathways. MCODE analysis showed two major gene clusters. 【Conclusion】 Apigenin mainly regulated cell proliferation, apoptosis, oxygen sensing and inflammation through PI3K-AKT, p53, HIF-1, NF-kappa B and other pathways, to further alleviate heat stress and hypoxia stress. The study provided a theoretical reference for the application of apigenin and apigenin-rich plants in dairy cow production.

Key words: apigenin, dairy cow, heat stress, hypoxia, network pharmacology, molecular docking