甲硫氨酸脑啡肽对猪前体脂肪细胞凋亡的调控机理
Regulation Mechanism of Apoptosis of Porcine(Sus scrofa)Precursor Adipocytes by Methionine Enkephalin
陈齐 1刘尊海 1任泽宇 1王文涛 1吕睿 1王思蒙 1孙超1
作者信息
- 1. 西北农林科技大学动物科技学院,杨凌 712100
- 折叠
摘要
甲硫氨酸脑啡肽(methionine enkephalin,Met-Enk)是一种阿片类神经肽,对于细胞增殖与代谢具有调控作用.为探究Met-Enk对猪(Sus scrofa)前体脂肪细胞凋亡及线粒体结构功能的影响,本研究使用Met-Enk处理猪原代前体脂肪细胞,利用细胞计数试剂盒(cell counting kit 8,CCK-8)、AV/PI(膜联蛋白V/碘化丙啶,annexin V/propidium iodide)染色技术检测细胞凋亡,通过透射电子显微镜(transmission electron microscope,TEM)和原子力显微镜(atomic force microscope,AFM)观察线粒体结构变化,并使用JC-1荧光探针等手段检测线粒体功能变化.结果表明,Met-Enk导致猪前体脂肪细胞活性下降,凋亡率升高,引起凋亡标记分子Caspase 3、Caspase 9等表达水平变化;TEM结果显示,前体脂肪细胞中线粒体数目减少、体积肿胀、线粒体嵴减少;AFM结果显示,线粒体发生形变,膜表面出现凹陷,粗糙度增加;结合免疫印迹和qPCR等方法发现,Met-Enk处理导致线粒体结构与功能的受损,并通过阿片生长因子受体(opioid growth factor receptor,Ogfr)促进Bax/Bak等凋亡因子表达,促使线粒体释放细胞色素C,促进细胞凋亡.本研究深入探讨了Met-Enk通过影响线粒体结构与功能诱导猪前体脂肪细胞凋亡的作用机理,为使用Met-Enk改善畜禽脂肪组织发育提供参考.
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.
关键词
甲硫氨酸脑啡肽(Met-Enk)/猪/前体脂肪细胞/细胞凋亡/原子力显微镜(AFM)Key words
Methionine enkephalin(Met-Enk)/Porcine/Precursor adipocytes/Apoptosis/Atomic force microscopy(AFM)引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据