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Regulation Mechanism of Apoptosis of Porcine (Sus scrofa) Precursor Adipocytes by Methionine Enkephalin |
CHEN Qi, LIU Zun-Hai, REN Ze-Yu, WANG Wen-Tao, LYU Rui, WANG Si-Meng, SUN Chao* |
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China |
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Abstract Methionine enkephalin (Met-Enk) is an opioid neuropeptide with regulatory effects on cell proliferation and metabolism. In order to investigate the effect of Met-Enk on apoptosis, mitochondrial structure and function of porcine (Sus scrofa) precursor adipocytes, in this study, Met-Enk was used to treat porcine primary precursor adipocytes, and detected apoptosis by using cell counting kit 8 (CCK-8) and annexin V/propidium iodide (AV/PI) staining, and observed mitochondrial structural changes by means of transmission electron microscope (TEM) and atomic force microscope (AFM) and detected the changes of mitochondrial function by JC-1 fluorescent probe and other methods. The results showed that Met-Enk treatment resulted in decreased activity and increased apoptosis rate of porcine precursor adipocytes, causing changes in the expression levels of apoptosis markers Caspase 3 and Caspase 9, etc. The TEM results showed that the number of mitochondria in the precursor adipocytes was reduced, the volume was swollen, and mitochondrial cristae were reduced. The AFM results showed that the mitochondria were deformed, and the surface of the membrane appeared to be concave and the roughness was increased; Immunoblot analysis and qPCR revealed that revealed that Met-Enk treatment led to impaired mitochondrial function and promoted apoptotic factors such as Bax/Bak through opioid growth factor receptor (Ogfr), which prompted the release of cytochrome C from mitochondria and promoted apoptosis. This study deeply investigated the mechanism of Met-Enk inducing apoptosis in porcine precursor adipocytes by affecting mitochondrial structure and function, so as to provide a reference for the use of Met-Enk to improve the development of adipose tissues in livestock and poultry.
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Received: 11 October 2023
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Corresponding Authors:
* sunchao2775@163.com
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