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本研究采用小鼠神经母细胞瘤无血清培养建立神经细胞老化实验研究模型。以显微分光光度仪测定的细胞内脂褐素荧光值和扫描电子显微镜观察的细胞表面结节状物的大小以及突出终末微突起数量作为实验性神经细胞的老化指标。结果发现:(1)脂褐素荧光值随细胞培养时间延长而增强;(2)细胞表面结节状物随增时而增大;(3)细胞微突起随增时而减少,最终消失。揭示细胞在无血清培养条件下由分裂增殖变为分化成熟老化,反映了神经细胞的老化过程。将脑啡肽加入培养液,结果如下:(1)使细胞内脂褐素荧光值显著减弱(P<0.01);(2)使细胞表面结节状物直径变小;(3)使细胞突起数量增加;④使细胞突起终末的微突起数量增加。结果提示脑啡肽可以延缓实验性神经细胞的老化进程。
In this study, mouse neuroblastoma serum-free culture to establish an experimental model of neuronal aging. The intracellular lipofuscin fluorescence measured by the micro-spectrophotometer, the size of cell surface nodules observed by scanning electron microscopy, and the number of prominent terminal microprotrusions were used as indicators of aging of experimental nerve cells. The results showed that: (1) Fluorescent value of lipofuscin increased with cell culture time prolonged; (2) cell surface nodules increased with increasing time; (3) cell microprojections decreased with time and disappeared eventually. Reveal the cells in the serum-free culture conditions from the proliferation of differentiation into mature aging, reflecting the aging process of nerve cells. Enkephalin was added to the culture medium and the results were as follows: (1) the intracellular lipofuscin fluorescence was significantly attenuated (P <0.01); (2) the diameter of the cell surface nodules became smaller; (3) The number increased; ④ the number of microprotrusions protruding terminal cells increased. The results suggest that enkephalin can delay the aging process of experimental nerve cells.