银杏叶提取物对MPP+诱导的中脑多巴胺能神经细胞损伤的保护作用

Protective effect of EGB761 on embryonic rat mesencephalic dopaminergic neurons in vitro from MPP^+-induced injury

目的:探讨银杏叶提取物(EGB761)对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病细胞模型是否具有保护作用.方法:采用胚胎大鼠中脑原代细胞培养法,分离培养中脑神经细胞,随后进行EGB761及MPP+处理,最后通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)微量比色法,酪氨酸羟化酶(TH)免疫细胞化学,研究EGB761对MPP+损伤后的中脑原代培养细胞及DA能神经细胞活性的影响,同时进行iNOS免疫细胞化学法,了解iNOS的表达情况.结果:在MPP+诱导的中脑原代细胞凋亡中,MPP+组TH阳性细胞数及细胞存活率均显著低于各EGB761组及阳性对照组(P<0.01).而iNOS的表达则显著多于各EGB761组(P<0.01).结论:EGB761对MPP+诱导的胚胎大鼠中脑原代培养细胞损伤具有保护作用,且此作用有随剂量的增大而增加的趋势.

AIM： To explore whether the ginkgo biloba extract （EGB761） could partially protect embryonic rat mesencephalic dopaminergic （DA-ergic） neurons from MPP^＋ -induced injury. METHODS： Primarily cultured cells were dissociated from embryonic rat mesencephalon and the cells were treated by addition of EGB761 and MPP^＋. The viability of mesencephalic primary cells and DA-ergic neurons was assessed by MTT assay and the survival number of tyrosine hydroxylase （ TH ） positive cells was evaluated by immunocytochemistry. The level of nitric oxide （NO） was assessed by immunocytochemistry for inducible nitric oxide synthase （iNOS）. RESULTS： The number of TH-positive cells and the viability of mesencephalic cells significantly decreased while the number of iNOS cells increased in the development of apoptosis induced by MPP^＋ . The adding of EGB761 significantly reversed these changes caused by MPP^＋. CONCLUSION： EGB761 can protect embryonic rat mesencephalic DA-ergic neurons against MPP^＋ -induced apoptotic death in a dose-dependent manner.