【摘要】 目的:研究胶质细胞源性神经营养因子(GDNF)和雌二醇(E2)联合使用对6-羟基多巴胺(6-OHDA)所致的黑质多巴胺能神经元损伤的保护作用及其机制。方法:新生2~3 d的雄性SD大鼠行常规中脑脑片培养,6-OHDA诱导损伤,脑片分为正常对照组、溶剂对照组、GDNF组、E2组、GDNF+E2组、PI3-K特异性阻断剂(wortmannin)+G1DNF组、wortmannin+E2组、wortmannin+GDNF+E2组。免疫印迹检测各组脑片中酪氨酸羟化酶(TH)、p-Akt、procaspase-3的表达。结果:在GDNF+E2组,TH的表达比单用GDNF或E2时显著升高,且procaspase-3的表达也比单用GDNF或E2时增高,差异均有统计学意义;GDNF、E2及GDNF+E2组中Akt磷酸化水平均是升高的,与相应的溶剂对照组相比差异有统计学意义;所有加入wortmannin组与相应的未加wortmannin各组相比,TH和procaspase-3的表达均是降低的,差异有统计学意义。结论:GDNF和E2联合使用对6-OHDA所致的黑质多巴胺能神经元损伤有保护作用,其机制可能是通过PI3K/Akt信号通路介导并且调节Akt下游的procaspase-3的表达变化实现的。更多还原

【Abstract】 Objective:To study the mechanism of GDNF and E2 on dopaminergic neuronal injury induced by 6-hydroxydopamine.Methods:The 2-3-day-old male SD rats were decapitated under sterile conditions,and then the slices were divided randomly into eight groups:normal control group,experimental control group,GDNF group,E2 group,GDNF plus E2 group,GDNF plus wortmannin(selective inhibitor of PI3-K) group,E2 plus wortmannin group,GDNF and E2 plus wortmannin group.The tyrosine hydroxylase(TH),p-Akt and procaspase-3 expressions in DA neurons were detected by Western blotting.Results:The expression of TH in GDNF in combination with E2 group was higher than that in the E2 and GDNF group while the difference was statistically significant(P<0.05).The expressions of procaspase-3 in GDNF and E2 groups were higher than those in GDNF or E2 group alone.The level of Akt phosphorylation was increased in GDNF,E2,and GDNF+E2 groups with a significant difference.The expressions of TH and procaspase-3 were decreased in the wortmannin groups,with a significant difference.Conclusion:GDNF and E2 have protective effects on dopaminergic neurons,mediated by P13-K/Akt pathway through regulating procaspase-3.更多还原