Action Mechanism of Rhubarb's Anti-cholestasis Based on Network Pharmacology and Experimental Validation
Objective To screen active ingredients of Rhubarb against cholestasis using network pharmacology,conduct an integrated analysis of its potential targets and mechanisms,and explore the possible action mechanism of Rhubarb through in vivo experiments.Methods The active ingredients and targets of rhubarb were retrieved and screened from TCMSP database,and the disease targets related to cholestasis were identified using GeneCards,OMIM,and DisGeNET databases.The regulatory network of"TCM-ingredient-target-disease"was constructed using Cytoscape 3.7.2 software,and GO and KEGG enrichment analyses were performed to predict the action mechanism of rhubarb against cholestasis.30 rats were randomly divided into 5 groups with 6 rats in each group:a normal control group(Control Group),an ANIT-induced cholestasis model group(75 mg·kg-1 ANIT administered by gavage,Model Group),a low-dose rhubarb group(75 mg·kg-1 ANIT+50 mg·kg-1 rhubarb water extract administered by gavage,Low Group),a medium-dose rhubarb group(75 mg·kg-1 ANIT+100 mg·kg-1 rhubarb water extract administered by gavage,Medium Group),and a high-dose rhubarb group(75 mg·kg-1 ANIT+150 mg·kg-1 rhubarb water extract administered by gavage,High Group).The effects of rhubarb water extract on serum biochemistry,liver damage,and protein expression in rats with cholestasis were observed.Results A total of 16 active ingredients and 56 related potential targets were screened,and 30 of the 2608 disease targets were found to be associated with rhubarb's anti-cholestasis.PPI network analysis identified TNF,TP53,CASP3,and IL1B(IL-1β)as potential core targets of rhubarb's anti-cholestasis action;GO and KEGG analyses revealed that the hepatitis B pathway,lipid metabolism and atherosclerosis,MAPK signal transduction pathway,p53 signaling pathway,and human immunodeficiency virus type 1 infection pathway were closely related to the anticholestasis effect of rhubarb.Rat experiments demonstrated that rhubarb water extract could improve serum biochemical abnormalities,significantly reduce the levels of TNFα and IL-1β(P＜0.05)in liver tissue,and decrease the protein expression of TP53,CASP3,and MYC in liver tissue(P＜0.05).Conclusion Rhubarb can achieve its action mechanism of anti-cholestasis by influencing target genes such as TNF,TP53,CASP3,IL1B(IL-1β),and MYC,as well as pathways including the hepatitis B pathway,lipid metabolism and atherosclerosis,MAPK signal transduction pathway,p53 signaling pathway,and human immunodeficiency virus type 1 infection pathway,by reducing inflammatory responses and improve cell apoptosis through the regulation of signaling pathways involving TP53 and Cleaved-CASP3.
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