论文部分内容阅读
目的探讨胶质细胞系源性神经营养因子(GDNF)促进中脑多巴胺(DA)能神经元存活和分化过程中,磷脂酰肌醇3激酶(P13K)信号通路和丝裂原活化蛋白激酶(MAPK)信号通路的可能作用。方法生后大鼠中脑脑片培养,并依培养基内加入的不同物质分为空白对照组、GDNF组、PI3K通路阻断组、MAPK通路阻断组。培养6 d后,进行酪氨酸羟化酶(TH)免疫组织化学染色,光镜观察,计算机辅助图像分析,统计学处理;同时用Westernblotting方法检测脑片中TH的表达。结果GDNF组TH表达阳性神经元的形状更趋向成熟,其TH表达阳性神经元的密度、胞体大小和TH的表达水平均显著高于空白对照组;P13K通路阻断组TH表达阳性神经元几乎消失,其TH表达水平显著低于GDNF组而与空白对照组无显著差别;MAPK通路阻断组TH表达阳性神经元的密度及TH表达水平与GDNF组无显著区别,但TH表达阳性神经元胞体显著小于GDNF组。结论P13K通路参与介导GDNF对DA能神经元的促存活作用,而MAPK通路参与介导其促形态学分化作用。
Objective To investigate the effects of glial cell line-derived neurotrophic factor (GDNF) on the survival and differentiation of dopaminergic neurons in mesencephalon. The effects of phosphatidylinositol 3-kinase (P13K) signaling pathway and mitogen-activated protein kinase ) The possible role of signaling pathways. Methods After birth, the rat brain slices were cultured and divided into blank control group, GDNF group, PI3K pathway block group and MAPK pathway block group according to different substances added into the culture medium. After cultured for 6 days, tyrosine hydroxylase (TH) immunohistochemical staining, light microscopy, computer aided image analysis and statistical analysis were performed. At the same time, the expression of TH in brain slices was detected by Western blotting. Results The neurons of TH positive neurons in GDNF group tended to mature. The density, cell body size and TH expression of TH positive neurons in GDNF group were significantly higher than those in blank control group. TH-positive neurons in P13K channel block group almost disappeared , The TH expression level was significantly lower than that of GDNF group and no significant difference with the blank control group; the density and TH expression level of TH-positive neurons in MAPK pathway block group were not significantly different from GDNF group, but TH positive cell bodies were significantly Less than GDNF group. Conclusions The P13K pathway is involved in the mediation of GDNF on the promotion of DA neurons. MAPK pathway is involved in the promotion of morphological differentiation.