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| Study on the mechanism of Kushen Wumei Decoction against hepatitis B virus in HepG 2.2.15 cell model based on network pharmacology and metabolomics |
| ZHENG Rui GUO Peng LIU Yu KONG Wei CHEN Yan |
| Department of Pharmacy, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing 100091, China |
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Abstract Objective To explore the anti-hepatitis B virus (HBV) mechanism of Kushen Wumei Decoction on HepG 2.2.15 cell model by using network pharmacology and metabonomics technology. Methods Enzyme linked immunosorbent assay was used to evaluate the effect of Kushen Wumei Decoction on HepG 2.2.15 cell model. Metabolomics analysis method of UHPLC-LTQ-Orbitrap-MS was used, combined with principal component analysis method and orthogonal-partial least squares multiplicative analysis method were used for multivariate statistical analysis. In addition, the network pharmacology technology was used to predict the potential metabolic pathways of Kushen Wumei Decoction against HBV. Combining the metabolic pathways obtained from the two parts, the anti-HBV mechanism of Kushen Wumei Decoction was explored. Results Compared with blank group, the survival rates of 1250 μg/ml and 2500 μg/ml groups were lower (P ＜ 0.05); Compared with blank group, 2500 μg/ml group had lower HBVDNA content (P ＜ 0.05); 40 metabolites with significant differences between groups were screened, which mainly affected the biosynthesis of phenylalanine, tyrosine, and tryptophan, phenylalanine metabolism, sphinolipid metabolism, arginine and proline metabolism; the potential therapeutic targets of the candidate active ingredients of Kushen Wumei Decoction were 115, and the top five targets in degree value were protein kinase B, human epidermal growth factor receptor, proto-oncogene tyrosine protein kinase Src, tumor necrosis factor, and vascular endothelial growth factor A. A total of 679 genes were identified by gene ontology enrichment analysis, including 2162 biological processes, 56 cell components, and 133 molecular functions; Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the top 20 P-value pathways mainly included PI3K-Akt, hepatitis B, Kaposi’s sarcoma associated herpes virus infection, and other signaling pathways. Conclusion Kushen Wumei Decoction can resist HBV. Metabolomics and network pharmacology found that the mechanism of its action may be related to the regulation of sphingolipid metabolism pathway, the biosynthesis of phenylalanine, tyrosine, and tryptophan, and phenylalanine metabolism pathway, and PI3K-Akt signaling pathway, which provides a basis for the mechanism of anti-HBV.
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