To Explore The Relationship Between Shugan Huazhi Capsules In The Treatment Of NAFLD And AMPK Signaling Pathway Based On Network Pharmacology

Purpose:Using modern network pharmacology technology,predict the targets and signal pathways of Shugan Huazhi Capsule in the treatment of chronic non-alcoholic fatty liver disease(NAFLD),and further analyze its mechanism of action.Materials and Methods: The active components and targets of seven Chinese medicines in Shugan Huazhi Capsule were searched through TCMSP database,ETCM database,BATMAN-TCM database,Pub Chem database,Swiss Target Prediction database and literature review.The Genecards database and OMIM database were used to retrieve and screen out the targets of NAFLD.The Uniprot database and Venny platform were used for the standardization and mapping of targets of Shugan Huazhi Capsule and NAFLD.And the network diagram of the main chemical components-targets of Shugan Huazhi Capsule was constructed by logging on the Cytoscape3.7.2 platform.PPI network analysis of drug target protein–disease target protein was performed using STRING database.The core targets were subjected to GO and KEGG enrichment analysis using DAVID database.Main enrichment pathway diagrams were obtained through visualization on Omishare cloud platform.Thirty healthy male rats of SPF grade were divided into three groups(10 rats in each group)by random number method.The rats in the normal group were fed with ordinary feed,while those in the model group and the ones soothing the liver and resolving stagnation were fed with high-fat diet for a feeding period of 56 days.After successful modeling was confirmed according to the diagnostic criteria of NAFLD,the required drug amount for rats was calculated,and the intragastric volume of rats was 1 m L/100 g.The rats of the other two groups were given normal saline by gavage,twice a day for 28 d.At the end of 28 d,the rats were sacrificed and the liver tissue was taken.Results:After screening,the active components of Chinese medicine were obtained:Rhizoma Pinelliae had 13 active components,Radix Bupleuri had 17 active components,Radix Salviae Miltiorrhizae had 65 active components,Semen Cassiae had 14 active components,Fructus Crataegi had 6 active components,Radix Curcumae had 15 active components,and Rhizoma Alismatis had 10 active components,totaling 140.The obtained247 targets of Shugan Huazhi Capsule and 1107 targets of non-alcoholic fatty liver disease were collected and screened.There were 103 intersecting targets of Shugan Huazhi Capsule and NAFLD.A network diagram of the main chemical components–targets was constructed on the Cytoscape3.7.2 platform.According to the degree value,the top ten key compounds were quercetin,luteolin,kaempferol,tanshinone,and baicalein.Through PPI network analysis,the protein-protein interaction network diagram was drawn,and the interaction proteins and core genes were obtained.GO enrichment analysis included cellular response to chemical stress,drug response,oxidative stress response,lipopolysaccharide response,and oxidative stress response in biological processes.KEGG enrichment analysis included AMPK signaling pathway,AGE-RAGE signaling pathway,IL-17 signaling pathway,tumor necrosis factor signaling pathway,and HIF-1 signaling pathway.Draw the target-pathway associated network diagram.The obtained core targets were AKT1,RELA,MAPK8,IKBKB,JUN,CHUK,TP53,CCND1,IL1 B,CDKN1A,CASP3,EGFR,GSK3 B,etc.The core pathways were AMPK signaling pathway,AGE-RAGE signaling pathway,IL-17 signaling pathway,and tumor necrosis factor signaling pathway.The results of confirmatory studies showed that compared with the normal group,the liver tissue SOD level and AMPK gene expression in the model group were decreased(P < 0.05).Compared with the model group,the SOD level and AMPK gene expression in the liver of the soothing liver and resolving stagnation group were increased,and the differences were statistically significant(P < 0.05).Conclusion:The mechanism of Shugan Huazhi Capsule may be related to AKT1,TP53,MAPK8 and other targets,and it can regulate AMPK,AGE-RAGE,IL-17,tumor necrosis factor,and HIF-1 and other signaling pathways to improve liver lipid metabolism,thereby inhibiting liver cell steatosis,so as to achieve the effect of treating NAFLD.Among them,AMPK is one of the core pathways in the treatment of NAFLD with Shugan Huazhi Capsule.SOD is the intersection target and one of the interaction proteins of Shugan Huazhi Capsule and NAFLD.Shugan Huazhi Capsule improved liver fatty degeneration by increasing the levels of AMPK and SOD,and played a role in the treatment of NAFLD.