| Glutathione S-Transferase(GSTs)is the second stage family of enzymes that are detoxified by the liver.It is mainly involved in the detoxification of various carcinogens,environmental toxins and oxidative stress products,which is the family of the a family of the GSTs superfamily A member who is also involved in the regulation of the signaling pathways caused by antioxidant stress and is also essential in the synthesis of prostaglandins.Broiler tibial cartilage dysplasia(TD)is the most common kind of leg disease in broilers,the main lesion in the growth plate.Ourprevious expression of chicken GSTA3 gene and obtain its protein activity;We use the results of the the application of gene chip technology,screened genes GSTA3,COX-2,PTGES,PTGDS,HPGDS,PTGR1,PTGER3,PTGER4 and related to the difference of the expression of prostaglandin synthesis.We investigated the effect of GSTA3 on the synthesis of prostaglandin in TD-induced broiler chickens,and then demonstrated the role of prostaglandin synthesis in broiler TD.Methods:according to the previous research method to construct the broiler TD model,and the prokaryotic expression of protein GSTA3 in different doses of intramuscular injection,the content of PGE2 ELISA in serum.TD of broiler tibial growth for quantitative study on the expression of GSTA3,COX2,PTGES,PTGDS,HPGDS,PTGR1,PTGER3,PTGER4,Ex-FABP gene by fluorescent plate quantitative PCR technology,through analysis and study of immunohistochemical expression of COX-2 protein.The results showed that HE staining,challenged group lesions and the pathological changes of TD,TD model was established successfully;injection of recombinant protein GSTA3,proliferative zone of cartilage cell morphology and arrangement in the later returned to normal,before the mast and mast cells still have lesions,but abnormal cells gradually reducedELISA results showed that in the early stage of thiram(experiment first,2,6 days)significantly increased serum levels of PGE2,the recombinant protein GSTA3 significantly reduced the level of PGE2 in the second and 4 days(P<0.05),on the fifteenth day of the experiment,the level of PGE2 is close to normal,the recombinant protein had no effect on PGE2.The results of quantitative PCR showed that the expression of GSTA3 was significantly down-regulated at 1,2,4 and 6 days after challenge(P<0.05).The expression of GSTA3 was not obvious after conteracting toxic substances.The expression of COX-2 gene in the test 1,2,4,6,and 10 days was significantly up-regulated(P<0.05).After injection of the protein,the expression of COX-2 was significantly lower in the low-dose group at day 1,2,4 and 6 days(P<0.05);HPGDS was significantly increased at 1,2,4 and 10 days(P<0.5),the same number of days were significantly reduced HPGDS(P<0.05)after injection of recombinant protein,and there was no significant difference in the expression of dose;The expression level of PTGDS was significantly increased after detoxification(P<0.05),and the expression of PTGDS was significantly down regulated(P<0.05),After injection of the protein the high dose of recombinant protein has obvious effect on PTGDS 0.05;The expression of PTGES was up-regulated in 1,4 and 10 days,and the expression of PTGES was decreased in the same day;The expression of PTGER3 was significantly decreased atl,6 and 15 days(P<0.05),after the injection of protein,the gene was significantly increased(P<0.05),and the low dose of recombinant protein was better;The expression of PTGER4 gene was significantly down-regulated on the 4th,10th,10th and 15th day(P<0.05).The expression of PTGER4 gene was significantly up-regulated at the same time(P<0.05)after the injection of protein,and the effect of high dose protein was better.PTGR1 did not change significantly in the test of 2nd,4th and 10th days,in 6th and 15th significantly decreased(P<0.05),injection of protein after high dose group were significantly up-regulated expression(P<0.05),low dose significantly decreased in fifteenth days(P<0.05),1st,2nd,4th,6th days no significant difference.The expression of Ex-FABP gene was significantly up-regulated(P<0.05),and the expression level of Ex-FABP gene was significantly decreased(P<0.05)There was no significant difference in the expression of dose group(P<0.05).Immunohistochemical results showed that COX-2 inoculation group in first days of positive signal enhancement,at the 2nd,4th,6th and 10th day,the positive signal weakened on the 15th day,after the injection of protein positive signal enhancement,atlst and 15th days,the positive signal weakened,the positive signal enhancement at the 2nd,4th and 6th day.The results showed that:1,TD occurred,the development process of PGE2 and related prostaglandin enzymes and receptors involved in growth plate chondrocytes abnormal proliferation may be related to prostaglandins;2,GSTA3 may be through the regulation of chondrocytes peanut tetraene Acid metabolism,prostaglandin synthesis pathway in the expression of related genes,inhibition of TD injury. |
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