Chronic Cold Stress Response Induced Nrf2Downstream Signal Pathway In Chicken

Chronic cold stress severely affect the growth and the production performance ofchicken through a complex physiological and biochemical reactions. So it is one of theimportant breeding target to enhance the ability of anti-cold stress in chicken. In this study,Huainan partridge chicken is experimental animal and its hepatocyte was cultured. Thehepatocyte was treated by KEAP1(Kelch-like ECH-associated protein1), the inhibitor ofNrf2(Nuclear factor erythroid-2p45-related factor2), and then it was used to study thetransduction of Nrf2antioxidant signal pathway through CHIP-Seq (Chromatinimmunoprecipitation sequence) and CoIP (Co-immunoprecipitation).1. Hepatocyte was got through perfusion, and it was cultured by adherent culturemethods in10cm diameter petri dishes. When hepatocyte cover the80%of petri dish thecells showed flat polygonal.2. The DNA enriched through CHIP-Seq and alignment showed that the sequence waslocated on chromosome1,2,3,4, W and Z. The binding site of transcription region, about12bp, was got through motif sequence analysis.3.10genes(VAT1L、TYRP1、TPH2、KCNAB1、HAAO、EFCBP1、DHRS11、CYP2H1、CYP1A4、ALDH1A1) in the antioxidant pathways, which were got through GO and KEGGPathway analysis.4. TPH2polyclonal antibody for chicken,was used in CoIP. The results of massspectrometry showed that TPH2mainly bind with fatty acid binding protein liver (127AA)probably in downstream. Fatty acid binding protein liver accounts for6.7%of the totalprotein.The results of the studies indicated that Nrf2antioxidant signal cascade was activatedin chronic cold stress in Huainan partridge chicken. The reaction pathway was probablyNrf2-TPH2-LFABP.