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To investigate the inhibitory effect and potential mechanism of Yinyanghuo (epimedii folium, EF) alcohol extract on hepatitis B virus (HBV) based on in vitro experiments and network pharmacology.
The wild-type HBV stably replicating cell line HepG 2.2.15 cells were treated with EF alcohol extract and the drug toxicity was evaluated using CCK8 assay. The HBV DNA, HBsAg and HBeAg levels were detected by PCR-fluorescent probe "one-tube method" and ELISA, and the inhibition rate was calculated. The TCMSP and GeneCards databases were used to screen the main active ingredients of EF and HBV-related targets, and the drug-active ingredient-potential disease target network was constructed with Cytoscape software. GO and KEGG enrichment analyses were performed on the core targets using the DAVID database.
The CC50 value of EF alcohol extract alone on HepG 2.2.15 cells was 1.21 mg·mL-1, and the maximum inhibition rates for HBV DNA, HBsAg and HBeAg were 59.54%, 63.07% and 45.73%, respectively (P<0.05). While the control drug tenofovir disoproxil fumarate (TDF) did not show significant inhibition for both HBsAg and HBeAg (inhibition rate <20.00%). The maximum inhibition rate of EF alcohol extract combined with TDF was 93.66% on HBV DNA, and the maximum inhibition rates on HBsAg and HBeAg were 54.97% and 61.28%, respectively (P<0.05). A total of 23 candidate active ingredients of EF and 104 potential targets for HBV treatment were screened by network pharmacology analysis. GO enrichment analysis yielded 624 entries, including 442 biological processes, 73 cellular components and 109 molecular functions; and the KEGG analysis mainly focused on signaling pathways such as cancer pathway, PI3K-Akt, lipids and atherosclerosis, human papillomavirus infection and hepatitis B.
EF can effectively inhibit the replication of wild-type HBV, and the combination of EF and TDF has a synergistic inhibitory effect, especially on antigen inhibition. EF has the characteristics of multi-component and multi-target action against HBV, which can be predicted to play roles through multiple signaling pathways. The findings provide a theoretical basis for subsequent in-depth exploration of the mechanism of EF in the treatment of HBV infection.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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