BDNF、NGF对体外长期培养的胚基底前脑胆碱能神经元的影响

本文探讨了脑源性神经营养因子、神经生长因子对体外长期培养的胚基底前脑胆碱能神经元是否具有延缓退变的作用。实验分为脑源性神经营养因子组、神经生长因子组、脑源性神经营养因子加神经生长因子组及单纯对照组。取孕 17d SD大鼠胚基底前脑原基制成细胞悬液接种于 2 4孔培养板中 ,按分组加入含相应神经营养因子和不含神经营养因子的 DMEM培养液 ,分别于体外培养 12、18、2 4、3 0 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 2 5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。结果显示 ,在培养的 4个时期 ,含脑源性神经营养因子组、神经生长因子组和脑源性神经营养因子加神经生长因子组的各项数据均明显地优于单纯对照组 ;脑源性神经营养因子加神经生长因子组的各项数据 ,特别是最长突起长度优于单独使用脑源性营养因子或神经生长因子组。提示 :脑源性神经营养因子和神经生长因子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大鼠胚基底前脑胆碱能神经元的退变 ;两者的联合使用还可对延缓其退变具有协同作用

EFFECTS OF BDNF AND NGF ON THE LONG-TERM CULTURED EMBRYONIC BASAL FOREBRAIN CHOLINERGIC NEURONS IN VITRO

The delaying effects of BDNF and NGF on the degeneration of the long term cultured embryonic basal forebrain cholinergic neurons in vitro were studied. The experiments were divided into BDNF, NGF, BDNF NGF and control groups. The cell suspension of embryonic basal forebrain was prepared from E17 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 five AChE positive neurons per well. The statistical data were treated by ANOVA and SNK test. In all four phases, the data of all groups containing BDNF or NGF were obviously superior to those of control group, and the data of BDNF NGF group, especially the length of the longest process, were better than those of group containing BDNF or NGF alone. These indicate that BDNF and NGF not only promote the development of the embryonic basal forebrain cholinergic neurons, but also delay the degeneration of the long term cultured embryonic basal forebrain cholinergic neurons in vitro; the combined use of two neurotrophic factors, BDNF and NGF, may have the synergic effects for delaying the neuronic degeneration.