Effects Of Recombinant GSTA3 And Ex-FABP Protein On Apoptosis-related And Prostaglandin-related Genes Expression In Erythrocytes Of TD Broiler

Tibial dyschondroplasia(TD)is a very popular group-occurring avian skeletal disease that is accompanied by apoptosis and inflammation.Prostaglandins(PGs)participate in bone formation and inflammation-related bone resorption in an autocrine/paracrine manner.Previously,we found that the expression of glutathione-S-transferase A3(GSTA3)and extracellular fatty acid-binding protein(Ex-FABP)have significant differences in the development of TD,and our research group has prepared the recombinant GSTA3 and Ex-FABP protein.In this study,the recombinant GSTA3 protein and recombinant Ex-FABP protein were injected separately into TD-induced broilers,and the effects of the two proteins on apoptosis-related and prostaglandin-related genes expression in erythrocytes were detected by real-time quantitative PCR and immunohistochemistry.The results of this experiment were as follows:(1)The transcripts of six apoptosis-related genes(Bcl-2,Bax,MDM2,BAG-1,BAG-3,and STAT3)and seven PGs-related genes(GSTA3,COX-2,PTGDS,PTGES,PTGER3,PTGER4 and PTGR1),and the protein expression of BAG-3,Bax and Bcl-2 were expressed in chicken erythrocytes.(2)Apoptosis-related genes expression were closely related to the occurrence of TD in erythrocytes,and the recombinant GSTA3 protein could promote the expression of all apoptosis-related genes in the early stage of TD and restored the normal expression of these genes in the TD recovery stage,which indicated that the recombinant GSTA3 protein could participate in the recovery of TD by suppressing the toxic effects of thiram.The immunoregulatory function of erythrocytes could be inhibited in the early stage of TD and promoted in the recovery phase by regulating the expression of PGs-related genes.In addition,the recombinant GSTA3 protein could participate in the recovery of TD by regulating the expression of PGs-related genes in erythrocytes.(3)TD could promote the apoptosis of chicken erythrocytes by suppressing the expression of anti-apoptotic genes(BAG-1,MDM2,STAT3),and also suppress nutrition transportation and the vascularization of cartilage.Further,the Ex-FABP protein did not significantly promote the expression of these apoptosis-related genes.The recombinant Ex-FABP protein did not significantly promote immune regulation in chicken erythrocytes in the early stages of TD.In the recovery stage of TD,chicken erythrocytes could restore the detoxification and PGE2 synthesis,which promoted PGs-related immune responses.Moreover,the mid-dose Ex-FABP protein could promote the expression of these genes and the recovery of TD better.In conclusion,this experiment demonstrated that there were many apoptosis-related and prostaglandin-related genes expressed in chicken erythrocytes,and the expression of each gene was closely related to the occurrence of TD.Recombinant GSTA3 protein could promote the anti-apoptotic and immunoregulatory effects of erythrocytes by regulating these genes expression.Recombinant Ex-FABP protein could participate in the immune regulation of erythrocytes in the recovery period of TD,but its anti-apoptosis effect was not obvious.