Proteomics On Key Regulatory Proteins And Regulatory Mechanisms Of Antioxidant Enzymes In Chicken Embryo Liver

During the development of the chicken embryo,the nutrients can only be provided by itself and it can not exchange substances with the world outside,and the metabolize waste must be cleared by itself.At the same time,the chicken embryo generates a large amount of active oxygen accumulated in the body as a result of the increased body metabolism during development,and high levels of active oxygen will cause irreversible damage to proteins,lipids and DNA.Some studies found that as the first antioxidant defense line,Peroxidase(POD),Superoxide Dismutase(SOD)and Glutathione peroxidase(GSH-Px)in chicken embryo work together to resist damage caused by reactive oxygen species.These antioxidant enzymes play an important role throughout the development of the chicken embryo,meanwhile the activity of the enzymes changes constantly.Therefore,it is very important to make clear the mechanism of antioxidant enzymes,but until now there has been little research in this area.Therefore,here further studied the liver tissues of chicken embryo eggs of Day12,Day16,and Day20 through proteome sequencing was carried out on basis of the research of the early transcriptome sequencing,and the key proteins that regulate antioxidant enzymes were identified during the development of chicken embryo to explore the regulation mechanism of antioxidant enzymes during chicken embryo development and the results were as follows,1: Identify the key proteins associated with the activity of antioxidant enzymes: the chicken embryo livers of Day12,Day16,Day20 were prepared to screen the differential proteins by the proteome sequencing,and the antioxidant enzyme activity differential proteins and enrichment pathways were screened through GO enrichment analysis and KEGG Pathway enrichment analysis.A total of 389 differential proteins and 50 enrichment pathways were found.The differential proteins related to antioxidants include: APOA1,APOA4,APOH,GPX1,GPX3,TTPA,HSP90AB1,etc.The KEGG Pathway related to antioxidants include: glutathione metabolism pathway,glyoxylic acid and dicarboxylic acid metabolism pathway and alanine,aspartic acid and glutamic acid metabolism pathway,etc.Comprehensive analysis confirmed that GPX1,GPX3,HAO2,TATP and other proteins were the key candidate proteins that affect the activity of antioxidant enzymes.2:Establish an oxidative model of chicken embryo liver cells: In order to study theanti-oxidation mechanism of liver cells,this study detected the effects of different concentrations of hydrogen peroxide on oxidase of chicken embryo liver cells by culturing chicken embryo liver cells.The effects of 2,4,6,8,10,and 12 mmol/L of hydrogen peroxide on hepatocytes were studied,respectively.The results showed that the optimal condition for the hydrogen peroxide chicken liver cell oxidation model was:hydrogen peroxide concentration of 8 mmol/L,Cultivate 4h.3: Taurine mediates the mechanism of GPX1's expression by regulating the anti-oxidation effect of chicken embryo liver cells: Taurine has obvious effect on anti-oxidation.In this study,the optimal concentration was chosen by adding different concentrations of taurine.Based on these results,the changes in the expression of GPX1 in the experimental group and the control group were identified to explore the mechanism of GPX1 regulating the anti-oxidation of chicken embryo liver cells.The results showed that taurine concentrations of 6.25 and 12.5 mmol/L can effectively inhibit the damage of chicken embryo liver cells by hydrogen peroxide.Quantitative PCR results showed that taurine can significantly increase the expression of GPX1,thus indicating that taurine may participate in the antioxidant effect of chicken embryos by mediating the expression of GPX1.This study is new to explore the antioxidant enzyme system of chicken embryo liver through proteomics technology.By establishing the oxidation model of chicken embryo liver cells,the role of taurine in inhibiting oxidative damage during the development of chicken embryo was detected,and its effect on the expression of GPX1 was also detected.This study provides a theoretical basis for further research on the development of chicken embryos and understanding the regulatory mechanism of embryo antioxidant enzymes during oxidative stress.