Genistein对体外培养的Parkinson病模型大鼠多巴胺能神经元及突触素的影响

为了探讨genistein(GST)对离体培养的Parkinson病模型大鼠多巴胺能神经元以及突触素的影响,取孕14~16d SD大鼠胚胎中脑腹侧部,常规体外培养。将培养的中脑腹侧细胞分为四组:正常对照组、雌二醇(E2)+MPP+组、GST+MPP+组、MPP+组。进行四甲基偶氮唑盐(MTT)法测定细胞活力和代谢状态。采用TH和突触素(synaptophysin,SYN)免疫荧光组织化学染色技术,观察各组TH阳性神经元的生长状况以及突触数量的变化。结果显示:与正常对照组、E2+MPP+组以及GST+MPP+组相比,MPP+组TH阳性神经元数量少,SYN免疫活性产物表达亦减少,细胞活力差。而E2+MPP+组、GST+MPP+组中上述指标与正常对照组相比,差异无统计学意义。以上结果提示Genistein对体外培养的多巴胺能神经元具有类似雌激素样的神经保护作用。

THE EFFECT OF GENISTEIN ON DOPAMINERGIC NEURONS AND SYNAPTOPHYSINS IN A MPP+ PARKINSON'S DISEASE MODEL IN VITRO

To study the effect of genistein on dopaminegic neurons and synaptophysin injured by 1-methyl-4-phenylpyridium ion (MPP~ ) in culture in vitro, the neurons from ventral mesencephalon of E14-16 SD rat were cultured and divided into four groups with different mediums: the control group only containing 10% fetal bovine serum in culture medium, E_2 MPP~ group, GST MPP~ group and MPP~ group. Methyl thiazolyl tetrazolium (MTT) was used to assay the cell viability and metabolism state. The cultured neurons were stained with immunofluorescence histochemical method by using antibody for tyrosine hydroxylase (TH) and synaptophysin (SYN) to observe the morphology and development of TH positive neurons and the change of synapses. The results showed that the number of TH positive neurons and SYN immunoreactive products in MPP~ group was low in comparison with those in control, E_2 MPP~ or GST MPP~ group (P<0.05). Meanwhile, the cell viability was also low in MPP~ group. No significant difference between control group, E_2 MPP~ group and GST MPP~ group were observed. These results indicate that genistein has neuroprotective effect similar to estrogen on dopaminergic neurons in vitro.