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.
Biochem Genet. 2022 Dec;60(6):2533-2551. doi: 10.1007/s10528-022-10233-4. Epub 2022 May 19.
Stanniocalcin-1 (STC-1) is a glycoprotein hormone involved in calcium/phosphorus metabolism and direct inhibition of bone and muscle growth. The aim of this study was to investigate the STC-1 gene with respect to the regulatory mechanisms of porcine growth metabolic pathways involving autophagy. Western blotting was used to detect the expression of autophagy and mitochondrial function-related proteins, and flow cytometry was used to detect mitochondrial function-related. Changes in the autophagosome and mitochondrial were observed by electron microscopy. The expression of the autophagy-related proteins was detected by confocal microscopy. The results showed that Pink1, Parkin and LC3B expression was increased; SQSTM1/P62 expression was reduced. Electron microscopy revealed that the cells in the serum starvation group all produced autophagosomes. The fluorescence intensity of GFP-LC3B and GFP-Parkin increased. The Bax/Bcl-2 ratio, Pink1 and Parkin protein levels were profoundly reduced in the STC-KO. In addition, the increase in Mfn2, OPA1, DRP1 and LC3B proteins was attenuated; the increase in the apoptosis rate and amount of active oxygen was attenuated; the decrease in membrane potential; the decrease in ATP was reversed; the fluorescence intensity of GFP-LC3B and GFP-Parkin was increased. These results indicate that autophagy can be caused by serum starvation. Knocking out the porcine STC-1 gene had an obvious antiapoptotic effect on cells, the inhibition of serum starvation-induced autophagy. This is the first study to show that the porcine STC-1 gene confers self-protection in the absence of nutrients. To provide a theoretical basis for studying the effect of STC-1 on pig growth and development.
钙结合蛋白 1（STC-1）是一种参与钙/磷代谢的糖蛋白激素，可直接抑制骨骼和肌肉生长。本研究旨在研究 STC-1 基因与涉及自噬的猪生长代谢途径的调控机制。Western blot 用于检测自噬和线粒体功能相关蛋白的表达，流式细胞术用于检测线粒体功能相关。通过电子显微镜观察自噬体和线粒体的变化。通过共聚焦显微镜检测自噬相关蛋白的表达。结果表明，Pink1、Parkin 和 LC3B 的表达增加；SQSTM1/P62 的表达减少。电镜显示，血清饥饿组的细胞均产生自噬体。GFP-LC3B 和 GFP-Parkin 的荧光强度增加。STC-KO 中的 Bax/Bcl-2 比值、Pink1 和 Parkin 蛋白水平显著降低。此外，Mfn2、OPA1、DRP1 和 LC3B 蛋白增加减少；细胞凋亡率和活性氧增加减少；膜电位降低；ATP 减少逆转；GFP-LC3B 和 GFP-Parkin 的荧光强度增加。这些结果表明，血清饥饿可以引起自噬。敲除猪 STC-1 基因对细胞具有明显的抗凋亡作用，抑制了血清饥饿诱导的自噬。这是首次表明猪 STC-1 基因在缺乏营养时具有自我保护作用的研究。为研究 STC-1 对猪生长发育的影响提供理论依据。
