Mechanism Of Gout In Gosling Induced By High Protein Diet

This study was designed to investigate the growth performance,blood biochemical parameters,liver and kidney tissue structure,expression of XDH and GLUT-9 in the liver and kidney,and the composition and quantity of cecal microflora of 1-14 days goslings fed high-protein diets,and to carry on preliminary discussion on the pathogenesis of gout.Seventy-two one-day-old geese were randomly divided into three groups.Group A,B and C were fed with diets containing 16%,20%and 24%crude protein,respectively.ELISA was used to detect inflammatory factors in liver and kidney tissues;liver and kidney tissues structure were observed by light microscopy and electron microscopy;expression of XDH and GLUT-9 in liver and kidney was detected by immunohistochemistry,real-time PCR and Western blotting;High throughput sequencing was performed to detect changes in the cecal microflora.The results are as follows:1.The growth performance of geese in group A were higher than those in group B and C.Serum total protein,albumin and globulin levels increased with the increase of dietary protein level,but the differences were not significant.Serum uric acid,alanine aminotransferase,urea nitrogen,and lactate dehydrogenase levels in group C were significantly higher than those in group A or B,while the total bilirubin levels in group C were significantly lower than those in group A and B.The interleukin-1β,interleukin-8 and xanthine oxidase activities of liver tissues and tumor necrosis factor-α activity of kidney tissues in group C were significantly higher than those in group A and B.2.H.E result displayed that the liver and kidney tissues were normal in group A;there were mild inflammatory cells in liver tissues and the kidney tissues were normal in group B;necrosis,edema and inflammatory infiltration of liver cells and tubular vacuolar degeneration,glomerular atrophy of kidney were observed in group C.Electron microscopy showed that the liver and kidney organelles were well-structured in group A;the mitochondrias were swelled and the mitochondrial cristaes were ruptured in liver and kidney cells of group B;the mitochondrias were vacuolar degeneration,glycogen was reduced,endoplasmic reticulum were swelled or disappeared in liver and kidney cells of group C.3.Immunohistochemical staining showed that XDH and GLUT-9 were mainly expressed in hepatocyte and the apical and basal sides of renal proximal tubular epithelial cell membrane.The absorbance values of XDH and GLUT-9 in liver and kidney of group C were extremely significant higher than group A(P<0.01).The results of real-time PCR showed that the relative expressions of XDH-mRNA and GLUT-9-mRNA in liver and kidney of group C were extremely significant higher than those in group A(P<0.01),which was significant higher than that in group B(P<0.05).Western blotting(Western-Blotting)showed that the relative expression of XDH and GLUT-9 protein in liver and kidney was significant higher in group B than in group A(P<0.05),and group C was extremely significant higher than group A(P<0.01).4.A Total of 1066175 high-quality bacterial 16S rDNA sequences were obtained from group A,B and C.The total number of OTU was 1013,and the representative contribution rate of principlecomponent 1(PC1)to total microbes was 46.64%,the contribution rate of principle component 2(PC2)was 16.46%.The dominant flora at the phylum level were Actinobacteria,Bacteroidetes,Firmicutes,Proteobacteria and Verrucomicrobia.The dominant flora at the family level were Bacteroidaceae,Enterobacteriaceae,Lachnospiraceae,Ruminococcacea,and Rikenellaceae.The dominant flora at the genus level were Akkermansia,Bacteroides,Corynebacterium and Lactococcus.Comparing with group A,the number of Bifidobacteria in group C was significantly lower(P<0.05),and the number of E.coli and Salmonella in group C were significantly higher(P<0.05).The experiment confirmed that,under the same feeding conditions,feeding the diet with 24%crude protein content affected the growth performance of the gosling and some blood biochemical indicators such as serum uric acid,affected liver and kidney function,caused liver and kidney tissue the damage,increased the expression of XDH and GLUT-9 of the liver and kidney,and the diversity and richness of the cecal microflora were changed.