Formula Le Cao Shi, a traditional Chinese medicine (TCM), has been used as folk remedy for treating hepatitis B for a long time. In our previous study, the anti-hepatitis B effects of Le Cao Shi have been verified. In the present study, the anti-hepatitis B activities of Le Cao Shi and its three single herbs were investigated in vitro by HepG2.2.15 cellular model, and the mechanisms against hepatitis B were deciphered via network pharmacology and gut microbiota analysis.
By network pharmacology method, twelve key compounds that played a vital role in Le Cao Shi were filtered from 213 ingredients. The targets RORA, CDK2, RELA, AKT1, IKBKG, PRKCβ and CASP3 were directly related to hepatitis B pathway, which indicated that Le Cao Shi could exert anti-hepatitis B effect by co-regulating cell cycle and inflammatory pathways.
The interactions between candidate compounds and target proteins that were directly involved in hepatitis B pathway were validated by molecular docking simulation and RT-PCR. By gut microbiota analysis, it was revealed that Le Cao Shi could alter the disordered microbial composition in the infected ducks towards normal, especially the restoration of three key strains, namely Streptococcus alactolyticus, Enterococcus cecorum and Bacteroides fragilis.
The above findings could provide a scientific basis for further development and utilization of Le Cao Shi against hepatitis B.