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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。
In order to investigate the protective effect of genistein (GST) on dopaminergic neurons in Parkinson model cultured in vitro, SD rat embryos were ventrally cultured in the ventral midbrain of 14-16 days and cultured in vitro routinely. The cultured cells were divided into four groups: normal control group, E2 + MPP + group, GST + MPP + group, MPP + group. Immunocytochemical staining was used to observe the growth of TH-positive neurons in culture. The staining of microtubule-associated protein-2 (MAP-2) in neurons was observed. Synaptophysin (SYN) immunofluorescence staining to observe the changes in the number of experimental groups of synapses. The results showed that compared with normal control group, E2 + MPP + group and GST + MPP + group, the number of TH positive neurons in MPP + group was decreased and the expression of SYN immunoreactive product was also decreased (P <0.05). The average gray level of MAP-2 positive staining Degree reduction of about 40%. However, there was no significant difference between E2 + MPP + group and GST + MPP + group (P> 0.05). The results of this study show that genistein has a similar estrogen-like neuroprotective effect on cultured dopaminergic neurons.