| This experiment investigated the potential immune-related genes of erythrocytes of broiler chickens in tibial dyschondroplasis（TD）,induced by thiram and genes related to TLRs signaling pathway.These immune-related genes regulate blood circulation and angiogenesis by stimulating erythrocytes,and this studied providing new insights into the therapeutic manipulation,for the treatment of TD prevention.In addition,immunity developed and TLRs signaling pathway related genes to assess the harmful effects of thiram,and to illuminate the molecular mechanism of immune-related genes related to TD.The regulatory starring role of Recombinant Glutathione-S-Transferase A3 Protein on immunerelated genes in TD-induced chickens were also investigated under this study.120,one day-old broilers were purchased and fed for 7 ds.After an overnight fast on the 7th d,all broilers were randomly divided into 6 groups A,B,C,D,E,and F.Groups D,E,and F were supplemented with 100 mg.kg-1thiram for 48 hours to induce the TD model.20 and 50 μg.kg-1rGSTA3 was added to groups B,C and E and F,respectively,to explore the therapeutic effect of rGSTA3 protein on TD chickens.Blood samples from each group of chickens were collected on d 6 and 10,and then sacrificed.Histopathology,hematoxylin and eosin（H&E）staining results showed that in the growth plate of TD broilers,the number,morphology,arrangement and distribution of chondrocytes were significantly changed,and TD symptoms were significantly aggravated on the 6th d with the addition of rGSTA3 protein TD symptoms eased in the group on d 10.The qRT-PCR results showed the expression of IRGs genes,most of which were upregulated in chicken erythrocytes at 10 ds.Ⅱ and other immune-related genes were upregulated on d 6,and p-38,MAPK12,CCL5,IRF8,IKBKE,KNB1 E,TLR2,4,5,6,7,21,LY6 E,LY86,LY96 and TRAF6 genes were significantly down-regulated on the same day.At the same time,immune genes related to the TLR signaling pathway were also evaluated in the TD chickens induced by thiram.CASP8,IRF3,5,8,10,AP-12,IKBKE,NFKBIE,NFKBIA,LY6 E,LY86,LY96,TLR1,2,3,4,21 genes were significantly up-regulated on day6（P < 0.05）and p-38,MAPK12,AP-2,MHC-I,TRAF6,IFN-γ,cMGF genes were downregulated on d 6.In addition,p-38,CCL-5,IRF5,8,NFKBIE,TLR1,4,7,IRAF6,IFN-γgenes were down-regulated on day 10.qRT-PCR results showed that rGSTA3 protein regulates immune-related gene expression,among which MAPK11,CASP8,IRF3,IRF10,AP-2,12,IKBKE were up-regulated on d 10（P < 0.05）.This study shows that injection of rGSTA3 protein in broilers can prevent the occurrence of TD in broilers,and the important immune-related genes IRF3,IRF5,IRF8,IRF10,TLR1,IRF2,IRF3,IRF4,IRF7,IRF21,MHC-1 and II,p-38.CCL5,AP-1,IKBK2,IKBK2,NFKBIE,NFKBIA,MAPK11 and 12,LY6 E,LY86,TRAF6,IFN-γ,cMGF have regulatory effects,potentially enhancing chondrocytes and erythrocytes in TD chickens by inducing TLR signaling pathway.Features.The findings suggest that rGSTA3 protein can treat broiler chicken TD induced by thiram by targeting immune-related genes,and provide a new prospect for poultry production. |
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