Metabolomics Was Used To Study The Mechanism Of QZRGF In The Prevention And Treatment Of Rat NAFLD

Objective:To explore the difference of liver metabolites between NAFLD rats and normal rats,the changes of liver metabolites after the effect of QZRGF,and to explore the mechanism of Quzhi Ruangan recipe in the prevention and treatment of NAFLD.Methods:A total of 96 SPF grade SD male rats were randomly divided into the control group,the model group,the Simvastatin group(1.8mg/kg),and the low,medium and high dose group of QZRGF(6.93mg/kg in the low group,13.86mg/kg in the middle group and27.72mg/kg in the high group),16 in each group.Except the control group,the other groups were fed high-fat diet for 12 weeks.Starting from the first day of the experiment,the rats were dissected at the end of the 8th and 12 th week respectively,the changes of TG,TC,LDL-C,HDL-C,AST,ALT and TBIL were observed.The degree of fatty change of liver tissue was observed by HE and Oil red O staining.At the end of 12 weeks,liver samples were prepared from control group,model group and QZRGF high dose group,detection of liver metabolites in three groups of rats by ultra high performance liquid chromatography tandem mass spectrometry(UHPLC-Q-TOF-MS),unsupervised principal component analysis(PCA)and supervised orthogonal partial least squares discriminant analysis(OPLS-DA)were used to process the data,Xcalibur software and Metlin were used to identify the differential metabolites,Metaboanalyst 3.0 was used to analyze metabolic pathways.Results:1.The rat model of fatty liver was established by feeding with high-fat diet,The liver index of model group was higher than that of normal group.At the end of 8th week,compared with the control group,lipid changes were observed in more than 1/3 cells per unit area of liver tissue in the model group,indicating successful modeling,disordered cell arrangement,blurred liver cords and mild inflammation in the HE staining.At the end of12 th week,the degree of lipid degeneration in the model group was worse than that at the end of 8th week,and a large number of fat vacuoles appeared in the liver cells,and the nuclei were extruded to one side of the cell membrane by slices of fat vacuoles,most of the hepatic cords were of unclear structure,with inflammatory cell infiltration.In the Oil red staining at the end of 8th week,a small amount of orange-red lipid drops appeared in the liver tissues of the model group compared with the control group,and most of the liver cords were still clear.At the end of 12 th week,compared with the normal group,the degree of hepatic steatosis in the model group was more severe than that at the end of 8 weeks,orange lipid droplets in liver cells were fused,and the structure of hepatic cords was blurred in most visual fields.2.The levels of TG,TC,LDL-C,AST,ALT and TBIL in serum of rats could be reduced by QZRGF.The content of HDL-C can be increased by QZRGF.3.In the mode of positive and negative ions,the substances in the control group vs model group can be divided into: organic acids,lipids and lipids,nucleotides,amines and vitamins,of which 11 substances are up regulated and 17 substances are down regulated.The pathways involved are taurine and hypotaurine metabolism,synthesis and degradation of ketone bodies,amimo sugar and nucleotide sugar metabolism,riboflavin metabolism,nicotinate and nicotinamide metabolism,glutathione metabolism,arginine and proline metabolism,glycine,serine and threonine metabolism.4.In the mode of positive and negative ions,the substances in the model group vs QZRGF high dose group can be divided into: organic acids,amines,nucleotides,heterocyclic compounds,sugars,lipids and lipids,vitamins,of which 19 substances are up regulated and 11 substances are down regulated.The pathways involved are taurine and hypotaurine metabolism,synthesis and degradation of ketone bodies,phenylalanine,tyrosine and tryptophan biology synthesis,glycerolipid metabolism,lysine degradation,cysteine and methionine metabolism,alanine,aspartate and glutamate metabolism.Conclusion:1.QZRGF can effectively prevent and treat nonalcoholic fatty liver disease induced by high-fat diet,reduce liver lipid deposition,reduce blood lipid and improve liver function.2.QZRGF can treat nonalcoholic fatty liver by strengthening spleen,activating blood circulation and softening liver,the metabolism of lipids,amino acids and vitamins in the liver changed,the results suggest that the mechanism of action may be related to synthesis and degradation of ketone bodies,glycerolipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,riboflavin metabolism.