PrP 106-126肽段对分化PC12细胞能量代谢的影响

Detection of Mitochondrial Membrane Potential and Activity of Ca^(2+)-ATPase of Differentiated PC12 Cell Infected by Prion Protein 106-126 Peptide

目的:研究朊蛋白106-126肽段对分化PC12细胞能量代谢的影响。方法:PC12细胞培养后加入神经生长因子(NGF),再加入朊蛋白106-126肽段,观察细胞存活率、线粒体形态学、线粒体膜电位和Ca2+-ATPase活性的改变。结果:细胞接触肽段后存活率下降28.9%;光镜下可见细胞贴壁不良,胞体缩小,细胞突起断裂缩短;电镜下可见核移位,染色质凝集,部分胞核呈碎块状,线粒体肿胀、嵴紊乱、断裂;膜电位和Ca2+-ATPase的活性随着肽段的作用时间的延长而减小。结论:朊蛋白106-126肽段对分化PC12细胞具有毒性作用,能量代谢障碍可能是其毒性作用机制之一。

Aim： To investigate the effects of energy metabolize of differentiated PC 12 cells after infected by prion protein 106-126 peptide. Methods： After differentiated by nerve growth factor （NGF）, the PC 12 cells were infected by prion protein 106-126 peptide. Cell viability was determined using the MTT assay and the morphological changes were observed. The mitochondrial membrane potential were detected by flow cytometry and the activity of Ca^2＋-ATPase were detected by spectrophotometer. Results： Infected by this peptide, cell viability decreased 28.9%. Under light microscope, cells were shrinked and rounded, many cells were divorced from plate wall, some neuraxon shortened and broke. Under electron microscope, chromatin gathered along the inside of the nuclear membrane, and mitochondria swelled and mitochondria ridge disconnected and disappeared. Along with the infected hours, decrease of mitochondrial membrane potential were detected by flow cytometry, and the activity of Ca^2＋-ATPase decreased were detected by spectrophotometer. Conclusion： Prion protein 106-126 peptide has cellular toxicity effect on differentiated PC 12 cells, and the energy metabolize disorder may be one of the reasons for the toxic mechanism.