The Effection Of Recombination GSTA3 And Ex-FABP On The Transcription Of Erythrocytes Immune Genes In TD Broilers Respectively

The transcription of some TLRs in erythrocytes have been demonstrated,the mRNA expression of other genes in erythrocytes were not clear.Tibial dyschondroplasia(TD)could result in the accumulation of many apoptotic chondrocytes.Previously,we demonstrated that chicken glutathione S-transferase A3(chGSTA3)and extracellular fatty acid-binding protein could contribute to the recovery of TD.In ordrer to explore the transcription of immune genes in erythrocytes,its influence to TD and the effection of GSTA3 and Ex-FABP on erythrocytes immune related genes respectively.Therefore,this study was to indentify the transcription of erythrocytes immune related genes,the transcriptional changes of immune related genes in TD broiler chickens and the effection of chGSTA3 and Ex-FABP on the transcription of these genes.respectively.The model of TD broiler chickens was established,in which 120 broiler chickens were randomly broken up into 6 groups;group A,B and C(fed with basal diet),group D,E and F(fed with basal diet containing 100 mg·kg-1 Thiram)after a week nonnal feeding.The broiler chickens of group B and E were treated with chGSTA3 protein by intramuscular injection(low dose:20 ?g·kg-1/bird)and the broilers of group C and F were treated intra-muscularly with chGSTA3 protein(high dose:50 ?g·kg-1/bird),while groups A and D received the same dosage of phosphate buffered solution(PBS).We used reverse transcription polymerase chain reaction(RT-PCR)to investigate the transcription of the 19 immune-related genes of chicken erythrocytes and we used real-time PCR to investigate the mRNA expression of the immune-related genes in chicken erythrocytes.The results demonstrated that TLR1,TLR2,TLR3,TLR4,TLR5,TLR6-(TLR1 B),TLR7,TLR15,TLR21,chicken interleukin-7(chIL-7),chicken interleukin-12p40(chIL-12p40),chicken interleukin-13(chIL-13),interferon-?(IFN-?),interferon-?(IFN-?),myeloid differentiation factor 88(MyD88),TNF receptor-associated factor 6(TRAF6),chicken myelomonocytic growth factor(cMGF),major histocompatibility complex class ?(MHCII),and major histocompatibility complex class ?(MHCI)were expressed in erythrocytes in mRNA level;contrasted with group A with phosphate buffered solution(PBS)treatment,the mRNA transcription level of TLR2,TLR3,TLR4,TLR5,TLR7,TLR15,MyD88,MHCII of Thiram-treated chicken erythrocytes in group D on the 4th day were significantly upregulated,whereas IL-7 was significantly downregulated;the transcription of immune-related genes:TLR2,TLR4,MyD88 in the chicken erythrocytes of group D were significantly downregulated than that in group A,but IL-7,TLR3,TLR5,TLR7,TLR15,MHCII were insignificant on the 15th day;The changes of above mentioned genes transcription in group E and F were significant except TLR4,TLR5,MyD88 in group E as compared with group D on the 4th day;The changes of TLR4,TLR5,MyD88 in group E and TLR3,TLR5,MyD88,IL-7 in group F were insignificant,whereas the others were significant compared with group D on the 15th day.Broiler chicken erythrocytes could induce the occurrence of TD in the early stage and alleviate TD symptoms in the late stage by changing the mRNA expression of IL-7,TLR2,TLR3,TLR4,TLR5,TLR7,TLR15,MyD88,MHCII.GSTA3 protein could play a role in TD by regulating the transcription of immune genes in erythrocytesAll broiler chickens were fed with basal diet for 7 days and then 120 broiler chickens were randomly broken up into 6 groups(group A,B,C,D,E and F),and 20 broilers in each group.Broilers in group A,B and C were fed with basal diet,whereas group A,B and C were given basal diet containing 100 mg·kg-1 Thiram.The broiler chickens in group B and D were treated intra-muscularly with 20 p?g·kg-1 Ex-FABP,group C and E received 50 ?g·kg-1 Ex-FABP,and group A and F were given phosphate buffered solution(PBS)at the same volume.The results showed that compared with group F with thiram treatment,the mRNA expression of TLR2,TLR3,TLR4,TLR5,TLR7,TLR15,MyD88,MHCII in group D and E were significantly downregulated on day 6 post-feed thiram,whereas IL-7 were significantly upregulated;the transcription of TLR2,TLR3,TLR4,TLR5,TLR7,TLR15,MyD88,MHCII in group D on day 15 post-feed thiram were significantly decreased than that in group F,and IL-7 were significantly increased;contrasted with group F,the transcription of TLR3 and IL-7 was unsignificant and the rest immune genes were significantly downregulated;The changes of TLR3,TLR5,MyD88,MHCII and IL-7 in group C and TLR2,7 in group B were insignificant,whereas the others were significant compared with group A on the 6th day;The changes of above mentioned genes transcription in group B and C were significant except IL-7 in group B as compared with group A on the 15h day.In conclusion,Ex-FABP could alleviate TD lesion by regulating the mRNA expression of TLR2,TLR3,TLR4,TLR5,TLR7,TLR15,MyD88,MHCII and IL-7.