Differential Proteomics Study On The Effect Of Sarmentosin On Fatty Liver In Tilapia

Purpose: To study the effect of different concentrations of Sarmentosin on fatty liver disease in tilapia,the expression and metabolic pathways of different proteins in each comparison group were analyzed by proteomics method,and the mechanism of Sarmentosin in the treatment of fatty liver disease was discussed,so as to provide theoretical basis for the application of Chinese herbal medicine in the treatment of fatty liver disease in fish.Methods: This experiment was divided into two stages.The first stage was the establishment of fatty liver model.The stage was divided into two groups,the low-fat group and the high-fat group.The successful establishment of the model was determined by measuring growth indexes such as body length and weight,liver body ratio,and HE staining.The second stage was the treatment stage for the addition of Sarmentosininto the feed.At this stage,the experimental fish were divided into six groups: NC group,FLSSB0 group,FLSSB50 group,FLSSB100 group,FLSSB150 group,and FLSSB200 group.Three replicates were set in each group,and ten fish were taken from each repeated group for protein extraction during sampling.The sample was labeled with i TRAQ,and the peptides were isolated by Thermo nano LC 1000 nm upgraded high performance liquid chromatography system,and identified by Q Exactive combined mass spectrometer.This experiment USES Proteome Discoverer 1.3software to submit the original map file of peptide identification to SEQUEST software for database retrieval.The expression ratio between the two groups was set to be greater than 1.2 or less than 0.83,and the p value less than 0.05 was the threshold for screening the differentially expressed proteins.The screened differentially expressed proteins were analyzed through KOGclassification annotation,KEGG Pathway enrichment analysis,KEGG Pathway enrichment analysis,differential protein expression pattern analysis,and differential protein network interaction analysis.Based on the above analysis results,KEGG pathway and key proteins related to the formation and treatment of fatty liver were screened.Results:1.Macroscopic observation showed that fishes in the high-fat group had slow swimming,enlarged abdomen,slow reaction and decreased appetite.After dissection,adipose deposit was observed in vivo,with white liver and yellow liver.After measuring and calculating the growth indexes,it was found that the weight of fish with fatty liver disease was lower than that of the normal group,but the ratio of liver to body was gradually higher than that of the normal group with the increase of weeks.The results of HE staining showed that the cells in the high-fat group were irregular in arrangement and the nuclei were severely deviated,which once again proved the successful establishment of the fatty liver model.2.The content of malondialdehyde,the physiological and biochemical indexes in serum was determined.The ALT content was the lowest when the dosage was 100 mg/kg,the AST content was the lowest when the dosage was150 mg/kg,the CAT content was the highest when the dosage was 100 mg/kg,and the MDA content was the lowest when the dosage was 150 mg/kg.3.According to KOGannotation and classification,when the dosage reaches150mg/kg,proteins related to lipid transport and metabolism function are significantly up-regulated,and the therapeutic effect on fatty liver is the most significant at this time.4.According to the KEGG Pathway analysis,high-fat diet will lead to down-regulated protein enrichment in the PPAR signaling Pathway,activate PPARα,PPARβ and PPARγ in adipose tissue,promote the down-regulated expression of related proteins,increase the accumulation of fat in the liver,and cause the occurrence of fatty liver disease.The addition of the drug will lead to the up-regulation of protein enrichment in the PPAR signaling pathway,which activates the PPARα,PPARβ and PPARγ in adipose tissue,promoting the up-regulation of related proteins and playing an obvious role in the treatment of fatty liver disease.5.A total of 334 differentially expressed proteins were screened by FLSSB150-vs-FLSSB0,including 199 up-regulated proteins and 135down-regulated proteins.These proteins are involved in ribosome cycling,PPAR signaling,methyl butyrate metabolism,linolenic acid metabolism,lipid metabolism,lipid binding,biosynthesis,citric acid cycling,AMPK signaling and other metabolic processes.Conclusion: through the observation,physiological index detection,bioinformatics analysis,and comparison of several groups of dosing groups,we judged that when the dosage of Sarmentosinwas 100-150 mg/kg,the therapeutic effect was the best.This study not only provides a new target for the treatment of fatty liver disease,but also has important significance for the production and development of efficient new Chinese herbal medicine.