神经生长因子和神经节苷脂GM1联合应用对鼠基底前脑胆碱能神经元的保护作用

Effects of combined NGF and ganglioside GMl on the embryonic basal forebrain cholinergic neurons in vitro

目的探讨神经生长因子(NGF)和神经节苷脂GMl联合应用对基底前脑胆碱能神经元是否具有延缓退变的作用。方法实验分为NGF组、NGF+神经节苷脂GMl组和单纯对照组。取孕20 d SD大鼠胚基底前脑原基制成细胞悬液接种于24孔培养板中,按分组加入NGF、NGF+神经节苷脂GMl和不含神经营养因子的DMEM培养液,分别于体外培养12、18、24、30 d后进行乙酰胆碱酯酶组织化学反应。显微镜下计数各孔中乙酰胆碱酯酶阳性神经元数,每孔随机测量和计数20个乙酰胆碱酯酶阳性神经元的平均胞体直径、发出的突起数和最长突起长度。数据用方差分析和SNK检验进行统计学处理。结果在培养的4个时期,NGF组和联合组的各项数据均明显优于单纯对照组,神经节苷脂GMl+NGF组的各项数据,特别是最长突起长度优于单独使用NGF组。结论NGF和神经节苷脂GMl联合应用及NGF单独应用不仅对体外培养的胚胆碱能神经元发育生长具有促进作用,而且还可延缓体外长期培养的人鼠胚基底前脑胆碱能神经元的退变;联合应用效果更佳。

Objective To investigate the effects of combined NGF and ganglioside GM1 on the degeneration of the long-term cultured embryonic basal forebrain cholinergic neurons in vitro. Methods The experiments were divided into NGF group, NGF ＋ ganglioside GM1 group, and control group. The cell suspension of embryonic basal forebrain was prepared from E20-day old SD rat. Dissociated cells were plated into the 24-well cultured plates which had DMEM medium containing corresponding neurotrophic factors and no neurotrophic factor according to the groups. The cultured neurons were detected by using AChE histochemical staining at 12, 18, 24, 30 days after culturing. The number of AChE positive neurons was counted and the average diameter of cell body, the number of processes and the length of the longest process were measured in twenty AChE positive neurons per well. The statistical data were treated by ANOVA and SNK test. Results In all four phases, the data of groups containing ganglioside GM1 or NGF were obviously superior to those of control group, and the data of ganglioside GM1 ＋ NGF group, especially length of the longest process, were better than those of NGF group. Conclusion These indicate that NGF ＋ ganglioside GM1, and NGF alone not only promote the development of the embryonic basal forebrain cholinergic neurons, but al so delay the degeneration of the long-term cultured embryonic basal forebrain cholinergic neurons in vitro.