Jiao Deling, Yang Zhen, Wang Lulu, Hu Binyue, Wang Jing, Xu Anyong, Cheng Wenmin, Jia Baoyu, Qing Yubo, Zhao Hong-Ye, Wei Hong-Jiang
Key Laboratory of Animal Gene Editing and Animal Cloning in Yunnan Province, Kunming, People's Republic of China.
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, People's Republic of China.
Anim Cells Syst (Seoul). 2019 Aug 16;23(5):318-325. doi: 10.1080/19768354.2019.1651766. eCollection 2019.
Leptin is an important adipokine and plays a vital role in animals. However, the role of leptin in the autophagic response of pig fibroblast cells (PFCs) has not been fully elucidated. In this study, we investigated the relationship between leptin and autophagy as well as underlying molecular basis. We found that PFCs treated with EBSS could secrete leptin, and the leptin concentration in the supernatant of leptin transgenic PFCs was higher than that of WT PFCs. We found an increase in LC3-II protein level and a decrease in p62 protein level in treated leptin transgenic PFCs compared with treated WT PFCs. Meanwhile, we observed an increase of autophagosomes by transmission electron microscopy and an enhancement of the accumulation of LC3 puncta in the cytoplasm of treated leptin transgenic PFCs, and these effects were further augmented by Baf A1 treatment. Furthermore, we detected the expression levels of 7 autophagy signaling pathway genes and 17 autophagy-related (ATG) genes by q-PCR. We found that between the two types of EBSS-treated cells 3 genes expression pattern were significantly different among the 7 autophagy signaling pathway genes and 8 genes expression pattern were significantly differernt among the ATG genes. These results indicated that leptin may promote autophagy and involving the downregulation of FOXO1 and LMNA genes via an unknown pathway which causes the upregulation of the 4 genes and the downregulation of 4 genes.
瘦素是一种重要的脂肪因子，在动物体内发挥着至关重要的作用。然而，瘦素在猪成纤维细胞（PFCs）自噬反应中的作用尚未完全阐明。在本研究中，我们调查了瘦素与自噬之间的关系以及潜在的分子基础。我们发现用EBSS处理的PFCs能够分泌瘦素，并且瘦素转基因PFCs上清液中的瘦素浓度高于野生型PFCs。我们发现与经处理的野生型PFCs相比，经处理的瘦素转基因PFCs中LC3-II蛋白水平升高，p62蛋白水平降低。同时，通过透射电子显微镜观察到自噬体增加，并且在经处理的瘦素转基因PFCs细胞质中LC3斑点的积累增强，而巴弗洛霉素A1处理进一步增强了这些效应。此外，我们通过q-PCR检测了7个自噬信号通路基因和17个自噬相关（ATG）基因的表达水平。我们发现，在两种经EBSS处理的细胞之间，7个自噬信号通路基因中有3个基因的表达模式存在显著差异，17个ATG基因中有8个基因的表达模式存在显著差异。这些结果表明，瘦素可能通过一条未知途径促进自噬，并涉及FOXO1和LMNA基因的下调，这导致4个基因上调和4个基因下调。
