College of Veterinary Medicine, Yangzhou University/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, Yangzhou 225009, Jiangsu, People's Republic of China; College of Animal Science and Technology, Jilin Agricultural University/Jilin Provincial Engineering Research Center of Animal Probiotics, Changchun 130118, Jilin, People's Republic of China.
College of Veterinary Medicine, Yangzhou University/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, Yangzhou 225009, Jiangsu, People's Republic of China.
Gen Comp Endocrinol. 2020 Jan 15;286:113298. doi: 10.1016/j.ygcen.2019.113298. Epub 2019 Oct 10.
Stanniocalcin (STC-1), a kind of glycoprotein hormone, was first found in fish and mainly regulates calcium/phosphorus metabolism in the body. To explore the biological function of the porcine STC-1 gene, the effects of changes in stanniocalcin expression on cellular metabolism and mitochondrial function were studied. A vector overexpressing the STC-1 gene and an siRNA silencer of the STC-1 gene were transfected into porcine kidney epithelial PK15 cells. After the STC-1 gene expression level was induced to change, STC-1 protein- and mitochondrial function-related proteins such as PMP70, OPA, DRP, Mfn and STC-1-related acetylated protein were detected by Western blotting. Cell apoptosis, mitochondrial membrane potential, reactive oxygen species (ROS), and ATP were detected using flow cytometry methods. Transmission electron microscopy was used to observe the changes in mitochondrial structure and morphology. The results showed that overexpression of the STC-1 gene could significantly upregulate the levels of PMP70, OPA, DRP and Mfn. STC-1 gene expression, which could decrease the apoptosis rate and reactive oxygen species production to significantly increase the cell membrane potential and reduce the formation of intracellular ATP, which also affected the morphology and number of mitochondria. The results were reversed when the STC-1 gene expression was silenced. The results suggested that the porcine STC-1 gene is closely related to cell growth metabolism and mitochondrial function, which influence the mitochondrial function-related proteins. The present study is useful for further understanding STC-1 gene function and provides a theoretical basis for improving the production characteristics of domestic pigs.
肌醇六磷酸（STC-1）是一种糖蛋白激素，最初在鱼类中发现，主要调节体内钙/磷代谢。为了探索猪 STC-1 基因的生物学功能，研究了 STC-1 表达变化对细胞代谢和线粒体功能的影响。转染过表达 STC-1 基因的载体和 STC-1 基因的 siRNA 沉默子到猪肾上皮 PK15 细胞中。在 STC-1 基因表达水平发生变化后，通过 Western blot 检测 STC-1 蛋白和与线粒体功能相关的蛋白，如 PMP70、OPA、DRP、Mfn 和 STC-1 相关的乙酰化蛋白。通过流式细胞术方法检测细胞凋亡、线粒体膜电位、活性氧（ROS）和 ATP。使用透射电子显微镜观察线粒体结构和形态的变化。结果表明，过表达 STC-1 基因可显著上调 PMP70、OPA、DRP 和 Mfn 的水平。STC-1 基因表达可降低细胞凋亡率和活性氧产生，显著增加细胞膜电位，减少细胞内 ATP 的形成，这也影响了线粒体的形态和数量。当 STC-1 基因表达被沉默时，结果则相反。结果表明，猪 STC-1 基因与细胞生长代谢和线粒体功能密切相关，影响线粒体功能相关蛋白。本研究有助于进一步了解 STC-1 基因的功能，为提高我国猪的生产特性提供理论基础。
