Study On The Intervention Mechanism Of Charred Angelica Sinensis Polysaccharide On Liver Injury Induced By Ceftiofur Sodium Combined With LPS In Chicken

Angelica sinensis,the dried roots of Angelica sinensis(Oliv.)Diels,is a kind of genuine traditional Chinese drugs in Gansu province.It has effects such as promoting blood circulation,nourishing blood,anti-inflammatory etc.The active components include volatile oil,organic acids,polysaccharides etc.Angelica sinensis polysaccharide has effect on regulating immunity and antioxidation.Charred Angelica sinensis(CAS)is one of the processed products of Angelica sinensis in clinical application.Polysaccharides from CAS(CASP)are the main functional components in charred Angelica sinensis.In this experiment,the mechanism of liver injury induced by ceftiofur sodium(CS)combined with LPS and the protective mechanism of CASP were studied by analyzing the changes of immune inflammatory factors and histopathology,combining with proteomics verification strategy of TMT and MRM.The experimental samples were divided into the normal control group,the model group,low dose intervention group of CASP(CASPL,0.125 g/kg/d)and high dose of intervention group of CASP(CASPH,0.25 g/kg/d),chicken serum and liver samples.In this study,the contents of IL-1,IL-4,IL-6,IL-10,TNF-? and TGF-? were detected by ELISA kit and pathological histology was examined.The chicken liver protein was detected and differential protein was screened by TMT technology,then verified by MRM technology.The results were as follows:1.The results showed that compared with the control group,contents of IL-1?,IL-6 and TNF-? were extremely significantly increased(P<0.01),significantly reduced(P<0.05)of IL-4 and TGF-?,extremely significantly reduced of IL-10(P<0.01)in the model group.Compared with the model group,contents of IL-1? and TNF-? were significantly decreased(P<0.05),extremely significantly decreased(P<0.01)of IL-6 and significantly increased(P<0.05)of IL-4 and TGF-? in the CASPH group,and contents of IL-1? and TNF-? were significantly decreased(P<0.05)and IL-10 decreased significantly(P<0.05)in the CASPL group.Compared with the control group,the model group had necrosis of hepatocytes,difficult identification of liver lobules,vacuolar degeneration and so on.Compared with the model group,the structure of liver cells in the casph group was more complete,and the degeneration and necrosis of liver cells in the CASPH group were significantly reduced,while the improvement of liver cells in the CASPL group was more obvious than that in the caspl group.2.A total of 6662 proteins were identified in the control group,model group and CASPH group.206 proteins were significantly different in Model/Control,among which 128 proteins were up-regulated and 78 proteins were down regulated.745 proteins were significantly different in CASPH/Model,among which 535 proteins were up-regulated and 210 proteins were down regulated.The differential proteins were concerned with related pathways such as sugar metabolism,lipid metabolism,amino acid metabolism,and inflammation,all of which were mainly enriched in immune related pathways.3.MRM technology was used to verify 19 differentially expressed proteins.The overall trend of TMT and MRM quantitative results was highly consistent.Seven differential proteins(gene names)(COX7A2L,FCER2,AGXT,CD34,GCAT,CYP2AC1,TBXAS1)were screened out,which were closely related to liver injury and liver protection mechanism,which may be marker proteins of CASP intervention mechanism of CS combined with LPS in chicken liver injury.In conclusion,the mechanism of CS combined with LPS was closely related to the immune inflammatory reaction,oxidative stress,amino acid metabolism and tissue repair.CASP alleviated degree of liver injury,regulated immune response and repaired liver injury.It may restore the abnormal condition of immune related inflammation and other important pathways by regulating inflammatory factors,the expression of FCER2 and TBXAS1 protein,up regulating expression of CD34?AGXT?GCAT?COX7A2L and CYP2AC1 protein in liver,which degree of oxidative stress injury caused by modeling factors,recovery of amino acid metabolism disorder and regulation of secretion of a variety of related enzymes can promote the repair of liver tissue and play the role of liver protection.