肌肉腌制过程中食盐介导PLIN1蛋白磷酸化调控ATGL和HSL水解肌内甘油三酯机制研究

肌内脂肪水解是干腌肉制品风味前体物形成的重要途径，也是肉品研究的重要领域。课题组前期研究发现腌制过程中肌内HSL活性显著升高、脂肪显著水解，但其具体机理还不清楚。因此，本项目拟以肌肉腌制为研究对象，借助电镜技术、磷酸化蛋白组学、Western blot、免疫荧光等技术手段，从组织、细胞、分子水平研究不同条件腌制过程中：1）肌肉微观结构、肌内脂肪分布以及脂肪细胞和脂滴形态变化规律；2）肌内脂肪细胞中PKA、AMPK、碱性磷酸酶活性及蛋白质整体磷酸化规律；3）脂肪细胞内PLIN1磷酸化对ATGL、HSL在细胞内转位及活性变化的影响；4）PLIN1磷酸化对ABHD5、G0S2与ATGL互作影响。通过分析这些变化与肌内甘油三酯水解以及腌制条件的相关性，阐明腌制过程中食盐诱导肌内脂滴形态变化以及PLIN1蛋白磷酸化调控ATGL、HSL水解甘油三酯机制，为干腌肉制品加工过程中脂质水解调控提供理论指导。

Intramuscular lipolysis is the important pathway of flavour precursors formation in dry-cured meat products, and is also the important research area of meat products. Our previous studies have found that the endogenous HSL activity was significantly increased and the fat hydrolyzed markedly. But it is not clear about its specific mechanism. So, the present project intends to take the muscle dry salting as research object, and carry out following research works from tissue, cellular and molecular levels by using the technical means of electron microscopy, phosphorylated proteomics, Western blot immunoblotting and immunofluorescence analysis etc. 1)the variation of muscle microstructure, intramuscular fat distribution and the morphology of intramuscular adipocytes and lipid droplets during the salting process of different conditions; 2)the changes of PKA, AMPK, alkaline phosphatase activity and overall protein phosphorylation in intramuscular adipocytes during the salting process of different conditions; 3)the effect of PLIN1 phosphorylation on the translocation and activity changes of ATGL and HSL in the intramuscular adipocytes during the salting process of different conditions; 4)the effect of PLIN1 phosphorylation on the interaction of ATGL with ABHD5 and G0S2 during the salting process of different conditions. Finally, by analyzing the correlation between these changes and intramuscular triglyceride hydrolysis and salting condition, the mechanism of salt induced intramuscular lipid droplet morphological changes and the regulation of PLIN1 phosphorylation on ATGL and HSL hydrolysis of intramuscular triglycerides were elucidated. This will provide theoretical guidance for the regulation of lipolysis in dry cured meat products during processing.