To Study Pharmacodynamics And Network Pharmacology Of Ganyusan Against Liver Fibrosis

Objective:To explore pharmacodynamics of the Traditional Chinese Medicine Compound Ganyusan in liver fibrosis.The possible pathway and the molecular mechanism of Ganyusan therapy for liver fibrosis were predicted.Methods:CCl₄ was injected into the abdominal cavity for 8 weeks and 12 weeks to establish animal models of liver fibrosis,respectively.Different doses of Ganyusan treatment was performed 15 day sequential therapy for 8 weeks liver fibrosis model and30 day sequential therapy for 12 weeks liver fibrosis model.Serum alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase were assayed with biochemical detection kits.Hematoxylin and eosin（HE）and Masson’s trichrome were evaluated liver injury and extent of collagen fibers.The level ofα-SMA was detected by immunohistochemistry and Western blotting.Information on Ganyusan bioactive compounds was collected from TCMSP and TCMID to identify the compound targets.Liver fibrosis–related targets were detected from Gene Card and OMIM database.The PPI network of Ganyusan-liver fibrosis was constructed on STRING database.The overlap targets of Ganyusan and liver fibrosis were identified for gene and pathway enrichment analysis by utilizing the GO and KEGG analysis in DAVID database.Using molecular docking analysis,the binding situation and interaction force of protein and small molecules can be predicted and obtained.The molecular structure was obtained from the Pub Chem database.The protein structure was obtained from the PDB database.Auto Dock Vina and Pymol were used in molecular docking.Results:The results of animal experiments showed that,in three treatment Ganyusan groups and Ganlixin group,serum levels of alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase were decreased than the model group.HE-stained tissue revealed that in the three doses Ganyusan treatment groups and the Ganlixin group inflammatory infiltration,hepatocyte swelling and necrosis,damage of the liver lobules were released compared with the model group.Masson staining showed less collagen in three treatment Ganyusan groups and Ganlixin group compared with the model group.Immunohistochemistry and Western blot results showed that the level ofα-SMA in three treatment Ganyusan group was lower than in model group.The results of network pharmacology showed that 90 compounds and 259targets were obtained from the TCMSP database and TCMID database.PPI protein network interaction diagram,drug-compound-target network diagram and compound-target-pathway network diagram were constructed respectively.Auto Dock Vina was used to conduct 3D docking between the active ingredients and the top 10 targets to obtain the docking results and display the closest renderings of the docking results.Conclusion:Ganyusan has a significant therapeutic effect on mice with liver fibrosis induced by CCl₄.The network pharmacology predicts that its possible key targets are PTGS2,PIK3CG,PRKACA,NOS2,AR,etc.The possible pathways include Pathways in cancer,Hepatitis B,Bladder cancer,TNF signaling pathway,Non-small cell lung cancer,etc.AR may be the most important target of Ganyusan in the treatment of liver fibrosis.