RNS/NO调控AMPK活化介导宰后牦牛肉线粒体细胞凋亡级联反应激活机制及对嫩度的影响

嫩度是评价肌肉品质和影响消费者购买欲的重要指标，也是制约我国牦牛肉精深加工和产业升级的关键影响因素。线粒体细胞凋亡途径被证实能改善肌肉嫩度，并受到NO和能量感受调节器AMPK二者相互作用的影响。NO是否通过调控AMPK活化介导线粒体细胞凋亡影响肌肉嫩化，相关研究尚未见报道。本项目主要研究NO调控AMPK磷酸化激活介导线粒体细胞凋亡发生机制及对牦牛肉嫩度的影响，通过建立肌肉原位模型和离体细胞模型，运用免疫荧光、免疫印迹、流式细胞术及iTRAQ磷酸化蛋白质组学技术探究NO水平变化调控AMPK磷酸化激活的具体位点和途径，以及AMPK活性调控对线粒体凋亡进程及肌肉嫩度的影响，揭示NO调控AMPK活化介导线粒体细胞凋亡级联反应发生机制及对牦牛肉嫩化的影响，为阐释RNS调控细胞凋亡影响宰后肌肉品质形成尤其对嫩度的改善提供理论依据，为深入阐述细胞凋亡嫩化机制和完善宰后肌肉成熟理论提供新的思路和理论指导。

Tenderness is an important index to evaluate meat quality and influence consumers' desire to buy and it is also a key factor to restrict the deep processing and industrial upgrading of yak meat in China. The mitochondrial apoptosis pathway has been shown to improve meat tenderness and is influenced by the interaction between NO and energy-sensing regulator named AMPK. But whether NO affects meat tenderness by regulating AMPK activity-mediated mitochondrial apoptosis has not been reported. This project mainly studies the mediation mechanisms of mitochondrial apoptosis cascade reaction and its effects on tenderizaion degree of yak meat by NO in regulating phosphorylation AMPK activation. The meat situ model and isolated cell model are estiblisted and immunofluorescence, western-blot, flow cytometry and iTRAQ phosphorylated proteomics are used to explore the NO level change in inluencing the regulation of phosphorylation AMPK activation of specific sites and ways, and the role of AMPK activity regulation on mitochondrial apoptosis process and meat tenderness. Finally, to reveal the mechanism of mitochondrial apoptosis cascade reaction and its influence on yak meat tenderization mediated by AMPK phosphorylation activatied by NO regulation. The results of this study will provide theoretical basis for elucidating the improvement of meat quality by RNS regulation apoptosis pathway especially the effects on meat tenderness. Furthermore, it is also help to expound the mechanism of apoptosis in regulating meat tenderization and provides new train of thought and theoretical guidance for improving meat tenderization theory during postmortem aging.