| Chicken Colibacillosis is a common and important infectious disease that endangers the health of chickens.In clinical practice,it is often prevented and treated with Ceftiofur sodium,which is easy to cause the accumulation of lipopolysaccharide(LPS)in chickens.Previous experiment studies about the drug intervention on chickens liver injury found that Ceftiofur sodium combined with LPS could cause obvious liver injury in chickens.The intervention effect on the liver injury of polysaccharides from Charred Angelica sinensis is accurate,but the mechanism is not clear.In this study,the samples collected from previous animal experiments about the drug intervention on chickens liver injury were taken as the research object,firstly the serum metabolomics study based on UPLC-Q/TOF-MS/MS technology was carried out.The histopathological observation was used for auxiliary analysis,and ELISA was used to verify the relevant metabolic pathways.Then the RNAref transcriptomic analysis of the chickens liver samples was carried out,and the correlation analysis with metabolomics was carried out.In addition,the detection of intestinal floras of chicken cecal contents by the 16 S r DNA amplicon sequencing technology and the short chain fatty acids detection of cecal content of chickens and their correlation analysis were carried out.The intervention mechanism of polysaccharides from Charred Angelica sinensis on liver injury induced by ceftiofur sodium combined with LPS was systematically analyzed from the perspective of metabolism,gene and intestine.The main results were as follows:1.The results of metabonomic studies showed that the typical metabolites closely related to liver injury included Deoxyinosine,L-Anserine,Uridine,Inosine,L-Lysine,(4Z,7Z,10 Z,13Z,16 Z,19Z)-4,7,10,13,16,19-Docosahexenoic acid,L-Glutamate,L-Leucine,1-Stearyl-2-oleoyl-sn-glycerol 3-phospholine(SOPC)and Succinate.The involved pathways mainly included Linoleic acid metabolism,m TOR signaling pathway and Amino acid metabolism pathway,and etc.The typical metabolites closely related to the hepatoprotective effect of polysaccharides from Charred Angelica sinensis included Ketoiscaproic acid,Ketoleucine,Taurine,L-Anserine,(4Z,7Z,10 Z,13Z,16 Z,19Z)-4,7,10,13,16,19-Docosahexenoic acid,L-Leucine,Deoxyinosine,Arachidonic acid,1-stearoyl-2-oleoyl-sn-glycerol 3-phospholine (SOPC).It was speculated that its hepatoprotective mechanism mainly involved Arachidonic acid metabolism,Linoleic acid metabolism,m TOR signaling pathway and Amino acid metabolism pathways.The results of histopathological observation showed that the chicken hepatocytes in the model group had severe necrosis,pyknosis of nuclei,extensive vacuolar degeneration and steatosis,expansion of most rough endoplasmic reticulum in cytoplasm,swelling of some mitochondria and autophagy.Chicken hepatocytes structure of the intervention group of polysaccharides from Charred Angelica sinensis was relatively complete,the hepatocyte degeneration was significantly reduced,a little autophagy in the cytoplasm,and the morphological structure wss close to that of the normal control group.ELISA showed that the liver protective mechanism of polysaccharides from Charred Angelica sinensis was indeed related to Arachidonic acid metabolism and m TOR signal pathway.2.The results of transcriptomic study showed that the liver protective mechanism of polysaccharides from Charred Angelica sinensis was mainly related to Arachidonic acid metabolism,m TOR signal pathway and amino acid metabolism pathway.The analysis of differential genes in Arachidonic acid metabolic pathway showed that compared with the normal control group,the expression of CLIP1 in the model group decreased significantly(P < 0.05),and the expression of ULK2 increased significantly(P < 0.05).The content of PLA2G4EL2 and the expression of CYP2J23 and CBR3 were significantly decreased in the intervention group(P <0.05).The analysis of differential genes in m TOR signal pathway showed that the expression of LOC107051846 in chickens liver of the intervention group of polysaccharides from Charred Angelica sinensis was significantly higher than that of the normal control group(P < 0.05).Compared with the model group,the expression levels of FZD4 and ULK1 in chickens liver of the intervention group of polysaccharides from Charred Angelica sinensis were significantly decreased(P <0.05).The correlation analysis between metabolomics and transcriptomics found that the screened differential metabolites had a strong correlation with the differentially expressed genes.Using the metabolites with R > 0.80 for pathway enrichment,it was found that the liver protecting mechanism of polysaccharides from Charred Angelica sinensis was indeed closely related to Amino acid metabolic pathway.3.The results of the study on the intestinal floras and short chain fatty acids found that compared with the normal control group,the intestinal floras in the model group was maladjusted.Compared with the model group,the ace,chao1,observed species and PD whole tree index of chicken cecum floras in the intervention group polysaccharides from Charred Angelica sinensis were significantly increased(P<0.05).It may be that polysaccharides from Charred Angelica sinensis can improve the micro ecology of chicken cecum floras,the diversity and richness of chicken intestinal floras changed significantly.It is speculated that the intervention mechanism of polysaccharides from Charred Angelica sinensis on chickens liver injury may be related to the abundance of some harmful and beneficial bacteria and the proportion of Bacteroidetes and Firmicutes.The correlation analysis results showed that Lactobacillus was positively correlated with the generation of valeric acid,acetic acid and butyric acid.Microbacterium and SMB53 were positively correlated with the generation of butyric acid,acetic acid,propionic acid and valeric acid.Granulicatella was positively correlated with the production of acetic acid and propionic acid,and Streptococcus was positively correlated with the production of propionic acid and butyric acid;Ruminococcus was negatively correlated with the production of isobutyric acid and valeric acid,faecalibacterium was negatively correlated with the production of valeric acid,and HA73 and W22 were negatively correlated with the production of propionic acid and butyric acid.In conclusion,this study confirmed that the intervention mechanism of the polysaccharides from charred Angelica sinensis on chickens liver injury induced by Ceftiofur sodium combined with LPS was closely related to Arachidonic acid metabolism,m TOR signal pathway,Amino acid metabolism pathway and the regulation of liver autophagy.Polysaccharides from Charred Angelica sinensis can improve the diversity of chicken cecal floras and affect the abundance and proportion of Bacteroidetes and Firmicutes.And it can interfere with the content of short chain fatty acids in chicken cecum by affecting the abundance of Lactobacillus,SMB53and Microbacterium.It is speculated that polysaccharides from Charred Angelica sinensis may interfere with chickens liver injury caused by Ceftiofur sodium combined with LPS by improving chicken cecal floras and affecting the content of short chain fatty acids.This study provided the theoretical foundation for screening veterinary medicine for liver protecting and ensuring chickens’health and food safety.This study is aim to provide basis for screening liver protecting drugs of chicken,protecting animal welfare and ensuring animal food safety. |
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