Genistein对体外培养的大鼠多巴胺能神经元的保护作用

The neuroprotective effects of genistein on dopaminergic neurons of rats in vitro

为了探讨genistein(GST)对离体培养的Parkinson模型多巴胺能神经元的保护作用,本实验取孕14～16 d的SD大鼠胚胎中脑腹侧,常规体外培养。将培养细胞分为四组:正常对照组、E2+MPP+组、GST+MPP+组、MPP+组。用免疫细胞化学染色法观察培养物中TH阳性神经元的生长状况、观察神经元中微管相关蛋白-2(MAP-2)的染色情况。通过突触素(SYN)免疫荧光染色,观察各实验组突触数量的变化。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免疫阳性产物表达亦减少(P<0.05);MAP-2阳性染色的平均灰度减少约40%左右。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(P>0.05)。本研究结果表明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。

To explore the neuroprotective effects of genistein on dopaminergic neurons 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, E2 ＋ MPP ^＋ group, GST ＋ MPP ^＋ group and MPP ＋ group. Using tyrosine hydroxylase （TH） immunocytochemical staining method to observe morphology and development of dopaminergic neurons, microtubule-associated protein-2 （ MAP-2 ） immunocytochemistry to show the dyeing status of neurons. The cultured neurons were stained with immunoflorescence by using antibody for synaptophysin （SYN） to observe the number change of synapses. The results showed that the number of the TH positive neurons and the SYN immunoreactive products in MPP ^＋ group were low in comparison with those in control, E2 ＋ MPP ^＋ or GST ＋ MPP ^＋ groups （P 〈 0.05）. Meanwhile, the average gray scale of MAP-2 positive staining in MPP ^＋ group reduced about 40%. However, no significant difference between control group, E2 ＋ MPP ＋ group and GST ＋ MPP ^＋ group were observed （ P 〉 0.05 ）. The present results suggest that genistein has neuroprotective effects similar to estrogen on dopaminergic neurons in vitro.