Mechanism Of Malondialdehyde On Autophagy And Apoptosis In Chicken During Maturation

In modern social life,commercial poultry has become the main food animal of human beings.Among them,chicken can provide animal protein with reasonable price,which is very important for human energy intake.Therefore,in the expansion of the chicken meat supply chain,research on the quality,quality and safety of chicken products,in particular,needs to be taken seriously.At present,most broilers on the market are slaughtered by cutting three tubes,which is prone to oxidative stress and induces lipid peroxidation.Therefore,the mechanism of the influence of a large amount of lipid peroxidation product malondialdehyde on the quality of post-mortem chicken is urgently needed.In this experiment,the yellow-footed broiler chicken in Yangling District of Shaanxi Province was used as the research object.The cell model was established by using the primary myoblasts of SD rats.The lipid peroxidation products were studied by Western blot,HE staining and cell fluorescence staining.The mechanism of the effects of malondialdehyde on autophagy,apoptosis and post-mortem maturation of chicken.The specific contents and results are as follows:1.To examine the effect of malondialdehyde on apoptosis,morphology and signaling pathway of myoblasts in SD rats.In this study,the primary SD rat myoblasts were used to simulate post-mortem hypoxia-ischemia as a cell model,and the concentrations and times of different malondialdehyde treatments were used to study the cell viability and morphology by MTT and AO/EB fluorescence staining.Optical microscopy;detection of oxidative stress in cells by ROS reactive oxygen species detection kit;endoplasmic reticulum stress(p-IRE1),mitochondrial apoptosis(Bax,Bcl-2,Caspase-9,Caspase-3)The expression of key factors of autophagy pathway(Beclin-1)and death receptor pathway(Fas,Fas-L)was detected by Western blot.The results showed that the expression of various signaling pathways was significantly increased(p<0.05)and the apoptosis was significantly increased at the lower concentration of malondialdehyde(0-0.1 mM).When the concentration was higher,the expression of each factor was significantly decreased(p<0.05),cell necrosis was significantly increased.2.Test the production of malondialdehyde in the mature process of broiler chickens after slaughter.In this study,the chicken breast meat of normal broilers was taken and stored in the refrigerator at 4 °C for 0,1,3,5 d,and the bilobalide acid method was used to detect the change of malondialdehyde content in chicken meat.The expression of Desmin in chicken was detected by Western blot.The results showed that with the prolongation of post-mortem time,the malondialdehyde content increased significantly(p<0.05);the Desmin expression intensity decreased significantly(p<0.05).3.Verify the effect of high concentration of malondialdehyde on the quality of post-mortem chicken.In this study,chicken breast meat was treated with 20 mM malondialdehyde,treated with physiological saline soaking as a control,Western blot was used to detect the expression of interstitial protein(Desmin);HE staining was used to detect changes in muscle fiber structure and muscle fiber gap;SDS-PAGE coagulation Gel electrophoresis was used to detect the basic skeletal structure changes of myofibrillar proteins.The results showed that the expression intensity of Desmin and the basic skeleton protein of myofibrillar protein were significantly decreased(p<0.05),indicating that the presence of high concentration of malondialdehyde was not conducive to the improvement of meat quality.4.Verify the effect of high concentration of malondialdehyde on the internal signaling pathway of post-mortem chicken.In this study,saline immersion treatment was used as a control.Chicken broth was treated with 20 mM malondialdehyde,major factors of autophagy pathway(Beclin-1,LC3-A and LC3-B),endoplasmic reticulum stress(BIP and The expression levels of IRE1),mitochondrial pathway-associated factors(Cleaved caspase-9,cyto-c)and apoptotic key protein(Caspase-3 zymogen)were detected by Western blot.It was found that the pathways in the malondialdehyde-treated group were not activated compared with the control group over time.The above results indicate that the chicken is matured with the production of malondialdehyde after slaughter;The lower concentration of malondialdehyde can activate the apoptotic signaling pathways in the cell model cells and promote the activation of Caspase-3;High concentration of malondialdehyde is not conducive to the improvement of meat quality during post-mortem maturation,and the cell autophagy and apoptosis pathways are not activated.