The Protective Effect And Mechanism Of Proteins In The Germinated Foxtail Millet(Setaria Italica) On Acute Liver Injury Induced By Carbon Tetrachloride In Mice

China has an abundance of millet resources,and foxtail millet contains a large amount of nutrients and has a high digestible portion.Germination can further enhance the nutritional value of grains.Currently,domestic and international studies on millet sprouts primarily concentrate on the whole grain powder or its bioactive components,like polyphenols and flavonoids,whereas studies on its protein are rather uncommon.The germinated foxtail millet protein has anti-inflammatory and antioxidant characteristics and may be helpful in the treatment and prevention of diseases brought on by oxidative stress and inflammation,possibly reducing liver damage.Therefore,this work enhanced the level of scientific cognition of the germinated foxtail millet protein and further provided the scientific basis for the development of germinated millet food by exploring the preventive impact of the germinated foxtail millet protein of acute liver injury by zoological studies.The main research methods,results and conclusions are as follows:1.As test animals,Balb/c mice were given low,medium,and high doses of germinated foxtail millet protein（LGR,MGR,and HGR）to treat acute liver damage brought on by a single injection of carbon tetrachloride（CCl₄）.To determine the germinated foxtail millet protein’s capacity to protect against the mice’s acute liver damage caused by CCl₄,several variables were analyzed,including the liver indexes,pathological sections,serum liver function indexes,and liver antioxidant indexes.The results showed that germinated foxtail millet protein reduced CCl₄-induced hepatic index abnormality,liver tissue injury,cell necrosis,and inflammatory cell infiltration.LGR,MGR and HGR significantly reduced the activities of AST and ALP in serum of mice with liver injury（P<0.01）.MGR increased the SOD activity in liver tissue,while HGR decreased the MDA content in.The findings demonstrated that the foxtail millet protein’s intervention improved the free radical scavenging ability of the liver,enhanced the antioxidant defense system of the body,and inhibited the lipid peroxidation,so as to play the role of liver protection.2.Total RNA was extracted from mouse liver tissue,and transcriptome sequencing（RNA-SEQ）was used to analyze the gene expression differences after HGR intervention.Compared with the Control group,4270 differential genes（2,249up-regulated and 2,021 down-regulated）were identified in the Model group.Compared with the Model group,229 differential genes（148 up-regulated and 81down-regulated）were identified in the HGR group.In the differential genes of CCl₄-induced liver injury mice interfered by HGR,genes related to liver disease were preliminarily screened,such as CX3CR1,LAIR-1,PTPRC and other significantly up-regulated genes,while CXCL3,GOT1 and other significantly down-regulated genes.3.GO enrichment analysis and KEGG enrichment analysis were performed for differential genes between Model group and HGR group.A total of 5,627 GO terms were obtained from GO enrichment analysis.Among them,the top 20 GO terms in the ranking of significance belong to biological processes,which mainly include response to external stimulus,system development,regulation of multicellular organismal process,and regulation of immune system process.221 pathways were identified from KEGG enrichment analysis.Differentially expressed genes were mainly involved in phenylalanine,tyrosine and tryptophan biosynthesis,cysteine and methionine metabolism,arginine biosynthesis,cytokine-cytokine receptor interaction,TGF-beta signaling pathway,Wnt signaling pathway,NF-kappa B signaling pathway,Fox O signaling pathway,chemokine signaling pathway,micro RNAs in cancer,and p53 signaling pathway.4.16S r RNA sequencing technology was used for high-throughput sequencing of intestinal microflora in the Control group,Model group and HGR group.The results show that the number of optimized sequences is 817,426 and the average length is 418.A total of 353 OTUs were obtained by clustering of the sequences of all samples,and all OTUs were annotated into 7 phyla,10 classes,30 orders,52 families,97 genera,and 144 species.There was no significant difference in theα-diversity index among all groups（P>0.05）,but theβ-diversity analysis showed that the microbial community composition was significantly different among different groups（PCo A:P=0.002）.At the phylum level,the relative abundance of Firmicutes increased and Proteobacteria decreased in HGR group.At the family level,the species with significant differences among the three groups were Rikenellaceae、Moraxellaceae、UCG-010,etc.At the genus level,the species with significant differences among the three groups were Alistipes、Acinetobacter、norank＿f＿＿Oscillospiraceae,etc.LEf Se analysis showed that norank＿f＿＿Oscillospiraceae,Monoglobales,Monoglobus and Monoglobaceae were rich in the intestinal tract of HGR group.In summary,germinated foxtail millet protein can protect the liver by enhancing the body’s antioxidant defense system,inhibiting lipid peroxidation,regulating the expression of chemokines and immune system-related genes,regulating amino acid metabolism and TGF-βsignaling pathway,and improving the composition of gut microbiota.