Differential Metabolic Pathways And Metabolites In A C57BL/6J Mouse Model Of Alcoholic Liver Disease And Preliminary Exploration Of Vitamin B6 Pathway

Objective:Explore the potential biomarkers of alcoholic liver disease and preliminary exploration of vitamin B6 pathway.Method:Acute alcohol model: C57 BL / 6 male mouses aged 5-6 weeks were randomly divided into a standard feed group(Pair-fed)and an alcohol-fed group(ETOH),5mouses in each group.After the mouses were fed for 10 days,the alcohol group used5 g / kg of mouse body weight as the standard,and 31.5% ethanol was administered to the stomach on the last day,while the mouses in the Pair-fed group were administered with an equal amount of dextrin solution.Nine hours after gavage,mouses were sacrificed,and serum was collected for blood biochemical indicators,including ALT,AST levels,and vitamin B6 and homocysteine levels;weighed after taking the liver;H & E and oil red O Hepatic lipid deposition was assessed by staining,and the remaining livers were quickly stored in liquid nitrogen for detection of liver TG content and metabolomics.Vitamin B6 supplementation model: C57 BL / 6 male mouse aged 5-6 weeks were randomly divided into a standard feed feeding group(Pair-fed)and an alcohol feeding group(ETOH),an alcohol plus vitamin B6 gavage group(ETOH + B6 gavage)Alcohol plus vitamin B6 feed group(ETOH + B6 feed)5 in each group.Nine hours after gavage,mouses were sacrificed,serum was collected and tested for ALT and AST levels;liver was weighed;H & E and oil red O staining were used to assess liver lipid deposition,and the remaining livers were quickly placed in liquid nitrogen Stored for the detection of liver TG content and for the extraction of total RNA.RT-PCR was used to detect liver inflammation-related factor levels and lipid synthesis-related gene expression.Results:In the model of the acute alcohol group,the weight of mice in the standard feed group showed an increasing trend.The serum serum alanine aminotransferase(ALT)and serum aspartate aminotransferase(AST)levels were significantly higher in the ETOH group than in the pair-fed group;liver weight,liver coefficient,and HE staining and oil red O staining in mice exposed to a single ethanol exposure revealed that the liver in the Et OH group had significant lipid deposition,and serum triglyceride(TG)levels were significantly increased,Nrf2 levels in mice exposed to a single ethanol were significantly lower than in the Pair-fed group;metabolomics analysis found three potential potentially different metabolic pathways in the positive ion channel(POS)and negative ion channel(NEG),respectively,the D-glutamine and D-glutamic acid metabolic pathways,the pyrimidine metabolic pathways in POS,Vitamin B6 metabolic pathway,as well as purine metabolism pathway,pentose phosphate pathway,cysteine and methionine metabolism pathway in NEG and related differential metabolites.In the model of the vitamin B6 supplement group,vitamin B6 in the alcohol group decreased,and homocysteine increased.The serum serum alanine aminotransferase(ALT)level in the ETOH group was significantly higher than in the pair-fed group.However,the vitamin B6 supplementation group significantly reduced the elevated serum alanine aminotransferase(ALT)levels and liver triglyceride(TG)content and lipid deposition caused by ethanol.The vitamin B6 supplementation group in the PCR significantly reduced the levels of inflammation-related factors IL1? and IL6 and expression of key liver lipid synthesis enzymes such as stearoyl coenzyme A desaturase-1(SCD-1)and acetyl coenzyme carboxylase(ACC).Conclusion:In the alcohol group model,six differential metabolic pathways with the most obvious changes and their potential biomarkers were mainly explored;it was found in the vitamin B6 supplementation model that VB6 can reduce the inflammation and lipid deposition caused by alcohol by blocking the lipid synthesis pathway.