Pink1介导的线粒体-内质网偶联调控猪肌内脂肪细胞分化的作用及机制

肌内脂肪含量是评价猪肉品质优良与否的关键指标之一，而线粒体的质量和功能完整性是调控肌内脂肪沉积的潜在要因。近来一项转录组测序发现线粒体自噬核心蛋白磷酸酶及张力蛋白同源物诱导激酶1(PTEN induced putative kinase 1, Pink1)与猪肌内脂肪的沉积能力高度相关。申请人前期研究发现，激活Pink1介导的线粒体自噬可显著抑制小鼠和猪皮下脂肪细胞的分化聚脂，提示Pink1是调控猪体脂沉积的重要基因。本项目拟通过体外培养猪肌内脂肪细胞，建立Pink1基因的干扰和过表达研究模型，利用透射电镜、免疫荧光和Seahorse细胞能量代谢等手段探究Pink1对猪肌内脂肪细胞分化聚脂的作用，采用CoIP-蛋白质组学技术筛选Pink1的潜在靶基因，从Pink1定位的线粒体与脂质合成的内质网偶联角度解析Pink1调控肌内脂肪细胞分化的潜在机制。本项目的实施将为猪肉质改良提供理论依据。

Intramuscular fat content is one of the key indicators to evaluate the quality of pork. The quality and functional integrity of adipocyte mitochondria are the key factors that regulate intramuscular fat deposition. A recent transcriptome sequencing study revealed that the mitophagy core protein Phosphatase and Tensin Homolog induced putative kinase 1 (PTEN induced putative kinase 1, Pink1) are highly correlated with the ability of intramuscular fat deposition in pigs. The applicant's previous research found that Pink1-mediated mitophagy significantly inhibits adipocyte differentiation in mice and pig, suggesting that Pink1 is an important gene regulating fat deposition of the pig. This project intends to establish a study model of Pink1 interference and overexpression by culturing porcine intramuscular adipocyte in vitro, and use transmission electron microscopy, immunofluorescence, seahorse approach and other methods to explore the role of Pink1 on the differentiation of porcine intramuscular adipocyte. The CoIP-proteomics was used to screen the potential target genes of Pink1, and the potential mechanism of Pink1 regulating intramuscular adipocyte differentiation through the coupling of the mitochondria targeted by Pink1 and the endoplasmic reticulum of lipid synthesis. The implementation of this project will provide a theoretical basis for pork quality improvement.