Study On The Mechanism Of Anti-liver Fibrosis Action Of Corydalis Saxicola Bunting Based On Liver Metabolomics And Network Pharmacology

Objectives:1.A metabonomics-based method was used to study the anti-liver fibrosis effect of Corydalis saxicola Bunting on CCl4-induced liver fibrosis in rats liver sample,thus revealing the mechanism of action of Corydalis saxicola Bunting in treating CC14-induced liver fibrosis in rats through selecting biomarkers and comparing the microscopic level of metabolic pathways with model group and treatment group.2.Using network pharmacology research methods to construct chemical composition-target-pathways-disease interaction network to find the effects of Corydalis saxicola Bunting CCl4-induced liver fibrosis in rat potential target.Further elucidate the mechanism of Corydalis saxicola Bunting in treating CC14-induced liver fibrosis in rat.3.Using molecular docking methods to elucidate CS compound interaction target with metabolomics and network pharmacology,aiming to find the effective components of CS in CCl4-induced liver fibrosis in rat.Methods:1.Thirty-two male Sprague-Dawley rats(180-220 g)were randomly divided into four groups:control group,model group,CS treatment group and colchicine treatment group.The model group,CS treatment group and colchicine treatment group received CCl4(0.1 ml/100 g body weight)and olive oil were mixed at the rate of 1:1(v/v)to induce hepatic fibrosis.The control group received olive oil(0.1 ml/100 g body weight,i.g.)and distilled water(0.5 ml/100 g,i.g.),the model group received CC14 olive oil solution(0.1 ml/100 g body weight,1:1,v/v,i.g.)and distilled water(0.5 ml/100 g,i.g.),the CS treatment group received CC14 olive oil solution(0.1 ml/100 g body weight,1:1,v/v,i.g.)and an i.g.dose of 2.5 g/kg body weight of CS(0.5 ml/100 g),and the colchicine group received CCl4 olive oil solution(0.1 ml/100 g body.weight,1:1,v/v,i.g.)and an i.g.dose of 0.1 mg/kg body weight of colchicine(0.5 ml/100 g).In addition,all of the groups except the control group were given an i.g dose of 0.1 ml/100 g body weight of 50%CC14 olive oil solution(0.1 ml/100 g body weight,1:1,v/v,i.g.)twice a week for ten weeks.All of the drugs were administered orally once a week begun at 7 weeks after modeling.2.UPLC-Q-TOF/MS metabonomics analysis techniques and multivariate statistical analysis were used to look for differences in liver metabolites between the model group and the control group at the metabolics level.To analyze the possible relationship between the differential substances and liver fibrosis,to identify potential biomarkers related to the treatment of liver fibrosis,and to further characterize the changes of the endogenous metabolites in the liver of model rats.The law found that CS interfered with the metabolic pathways of liver fibrosis and explained the mechanism of CS in the intervention of liver fibrosis.3.Searching compound of CS through data and literature,based on the compounds of CS to predicted targets of liver fibrosis.4.Query the relevant gene targets of liver fibrosis through the database.5.Query the relevant gene targets of metabolism pathway by CS regulate through the database.6.Establish network of CS-chemical-target-disease-metabolism pathway relationship using network visualization software Cytoscape 3.2.1 software to find related gene targets.7.Integrate the results of metabolomics and network pharmacology to discuss the specific intervention mechanism of CS on liver fibrosis.8.The Sybyl 2.0 Molecular Docking Software and the SystemsDock Online Molecular Docking Site were used to explore the effective chemical composition against liver fibrosis of CS.Results:1.Six potential biomarkers were found in the liver of CC14-induced liver fibrosis in rats,in which the levels of Oxidized glutathione and N-Acryloylglycine were elevated,and the levels of phosphorylcholine,uric acid,riboflavin and pantothenic acid were decreased.It mainly affects pantothenic acid and CoA biosynthesis,glutathione metabolism,glycerolphospholipid metabolism and purine metabolism pathways.2.CS can down-regulate phosphorylcholine,Oxidized glutathione and N-Acryloylglycine in the liver of liver fibrosis in rats and up-regulate uric acid,riboflavin and pantothenic acid.It is mainly responsible for the metabolism of pantothenic acid and CoA biosynthesis,glutathione metabolism and purine metabolism caused by CCl4-induced liver fibrosis in rat.3.The network pharmacology found that there were 45 active ingredients in CS and 264 diseases related to liver fibrosis.A total of 8 related gene targets were found after using the Cytoscape software to establish the CS--chemical-target-disease-metabolism pathway network.4.The molecular docking found that Metabonomics,network pharmacology and.molecular docking analysis showed chelerythrine,sanguinarine,cavidine,dehydrocavidine and feruloylagmatine are potential active compounds of CS to against liver fibrosis after CCl4 induction.Conclusions:1.The CCl4-induced liver fibrosis rat model can cause a significant change in the metabolic profiles of the pantothenic acid and CoA biosynthesis,glutathione metabolism,glycerolphospholipid metabolism and purine metabolism pathways.CS has a corrective effect on pantothenic acid and CoA biosynthesis,glutathione metabolism and purine metabolism.2.Metabonomics,network pharmacology and molecular docking analysis showed CS anti-liver fibrosis in rat through regulate the targets of ALT,FXR,COX-2,MMP-1,AGT,GGT1,FHIT and GPD1.Chelerythrine,sanguinarine,betulinic acid,?-amyrin acetate and pallidine are potential active compounds of CS anti-liver fibrosis through regulate targets of ALT,FXR,COX-2 and MMP-1.3.Chelerythrine,sanguinarine,cavidine,dehydrocavidine and feruloylagmatine are potential active compounds of CS to cure liver fibrosis after CCl4 induction.