Genistein对大鼠基底前脑胆碱能神经元发育影响的体外研究

Effect of genistein on the development of the rat basal forebrain cholinergic neurons in vitro

本文旨在研究染料木素(genistein)对体外培养的基底前脑胆碱能神经元发育的影响。取孕18d胎鼠基底前脑神经元,体外有血清培养7d后,随机分为3组:无血清培养液组(control组),genistein+无血清培养液组(G组)、E2+无血清培养液组(E2组)。48h后用MTT法测定细胞活力和代谢状态;用AChE组化染色以及ChAT和MAP2免疫荧光双重染色,镜下计数AChE阳性和ChAT阳性神经元数,并对两种神经元的细胞面积、第一级突起数及最长突起长度进行检测。结果显示:MTT法所测的OD值、AChE阳性和ChAT阳性神经元数、细胞面积、第一级突起数及最长突起长度在G组与control组之间无明显差异,然而E2组中的上述数值均明显增加。本研究的结果提示:genistein对体外培养的基底前脑胆碱能神经元无类似雌激素样的神经保护和营养作用。

To study the effects of genistein on the development of the rat basal forebrain eholinergie neurons. Neurons from the rat basal forebrain of embryonic 18 d were cultured for 7 d, and then divided randomly into 3 groups. Control group： the culture medium contained no serum. G group： the genistein was added into the serum-free culture medium. E2 group： 17β-estradiol was added into the serum-free culture medium. Two days later, the cell viability and metabolic state of the neurons were assayed by MTT method, and the cultured cholinergic neurons were detected by AChE histochemistry and ChAT / MAP2 immunofluorescence double-staining. The numbers of AChE and ChAT positive neurons were counted under microscope, the cell areas and the number of the primary processes and the length of the longest process were analyzed with image analysis system. The results showed that between the G group and control group, there were no significant difference in OD value of the MTT, the number of the AChE and ChAT positive neurons, the cell areas, the number of the primary processes as well as the length of the longest process. However, these above data were significantly higher in E2 group than these in the control group and G group. These results suggest that genistein has no neuroprotective or neurotrophic effect as that estrogen has on basal forebrain cholinergic neurons in vitro.