Establishment Of Liver Fibrosis Mouse Models And Screening For Serum Biomarkers

Background and Purpose: Liver fibrosis refers to a common pathological manifestation of various chronic liver diseases and is an important process in the diagnosis and treatment of liver diseases.Currently,clinical diagnosis of liver fibrosis mainly relies on pathology,imaging and laboratory tests,but more accurate and effective diagnostic methods are still needed.Furthermore,due to the complex pathogenesis of liver fibrosis,there are currently no drugs with clear curative effects available.Before clinical research,it is necessary to establish liver fibrosis mouse models that can simulate authentic liver fibrosis caused by different etiologies to prevent and treat liver fibrosis in basic research.Metabolomics is an integral part of systems biology,which is used to explore the pathogenesis of diseases.Biomarkers are the research significance of metabolomics.Considering the lack of effective diagnostic biomarkers and drug treatments in clinical practice,screening for the serum biomarkers of liver fibrosis models through metabolomics can not only provide a preliminary theoretical basis for clinical metabolomics,but also have important biological significance for the study of metabolic pathways.This project aims to analyze the serum metabolites of different liver fibrosis mouse models through metabolomics and to find common metabolic markers and metabolic mechanisms related to the occurrence and development of liver fibrosis.Methods: Using chemical inducer CCl4,high-fat and high-sucrose(HFHS)diets and bile acid ligation,five different mouse models were established and verified by serum biochemical indicators,liver pathological analysis and fibrosis markers.The serum samples of the above mouse models were detected by UPLC-HDMS technology.The differential compounds were screened by multivariate statistical analysis methods,and then matched with the metabolomics databases to identify specific endogenous metabolites of each models.Pathway analysis was used to study the changes in metabolic pathways related to liver fibrosis.Finally,the differential metabolite information and metabolic pathways of the five liver fibrosis models were integrated.Some common liver fibrosis biomarkers were screened.Results: In the five liver fibrosis models,serum AST and ALT levels of the Model groups were increased differently than those of the Control groups,but they were not increased significantly in the HFHS diets-induced early liver fibrosis model.H&E staining of liver tissue sections found that inflammation and liver injury occurred in the Model group in the five models.In terms of liver fibrosis evaluation,Masson staining and Picrosirius Red staining showed that the Model groups in the five models had different degrees of collagen fiber deposition and the severity of liver fibrosis was different.The m RNA expression levels of Acta2 and Col1a1 encoding liver fibrosis genes further verified the pathological results,which were both increased in the Model groups.The above results indicated that five liver fibrosis mouse models had been successfully constructed.Then,a total of 81 differential metabolites of the CCl4 acute model,63 differential metabolites of the CCl4 chronic model,88 differential metabolites of the HFHS early model,107 differential metabolites of the HFHS advanced model and 147 differential metabolites of the cholestatic model were screened by multivariate statistical analysis.Metabolic pathway analysis revealed that the pathways related to liver fibrosis were phenylalanine metabolism,tryptophan metabolism,arachidonic acid metabolism,glycerophospholipid metabolism,retinol metabolism and biotin metabolism.The integrated analysis of the differential metabolites of five liver fibrosis models found that the common metabolic markers are Lyso PC(14:0),Lyso PC(15:0),Lyso PC(16:0),Lyso PC(17:0),Lyso PC(18:0),Lyso PE(22:0/0:0),12(S)-HEPE and 4,7,10,13-Eicosatetraenoic acid.Conclusions: Different types of liver fibrosis mouse models can be replicated successfully.The metabolic pathway changes involved in the occurrence of liver fibrosis included phenylalanine metabolism,tryptophan metabolism,arachidonic acid metabolism,glycerophospholipid metabolism,retinol metabolism and biotin metabolism.Lyso PC(14:0),Lyso PC(15:0),Lyso PC(16:0),Lyso PC(17:0),Lyso PC(18:0),Lyso PE(22:0/0:0),12(S)-HEPE and 4,7,10,13-Eicosatetraenoic acid can be used as serum biomarkers of liver fibrosis.