银杏内酯B对胚胎大鼠中脑神经细胞的作用研究

目的:研究银杏内酯B(GBE)促进和保护胚胎大鼠中脑神经细胞的作用。方法:采用14d胚胎大鼠中脑神经细胞原代培养,实验分为7组,即正常对照、GBE对照(50mg·L-1)、海人藻酸(KA,100 μmo·lL-1)、神经生长因子(NGF,50mg·L-1+KA100 μmol·L-1)和GBE高、中、低剂量(100、50、25mg·L-1+KA100 μmol·L-1)组。分别培养胚胎大鼠中脑神经细胞10d,NSE免疫组化法染色鉴定神经元;硝酸还原酶法测定一氧化氮(NO)含量;改良的硫代巴比妥酸法测定丙二醛(MDA)含量;黄嘌呤氧化酶法测定超氧化物岐化酶(SOD)活性。结果:GBE高、中、低剂量组细胞生长、分化程度明显好于KA组。与正常对照组比较,KA组SOD活性显著减弱,MDA、NO含量显著增加;与KA组比较,GBE高、中、低剂量组SOD活性显著增强,MDA、NO含量显著减少。结论:GBE可促进中脑神经细胞生长与分化,对抗KA毒性作用,其机制可能与GBE减轻细胞脂质过氧化、抗自由基作用有关。

Effects of Ginkgolide B on Embryonic Rat Midbrain Neurons

OBJECTIVE： To study the promotion and protective effects of ginkgolide B （GBE） on the growth and differentiation of embryonic rat midbrain neurons. METHODS： The primary cell culture was made in vitro by using midbrain neuronic cell suspension from 14 days old Wistar rats. Rats were divided into normal control group, GBE control（50 mg.L-1） group, KA（100μmol·L-1）, NGF （50 mg. L I＋KA100 μmol. L-1）, GB high-dose, medium-dose and low-dose （ 100,50,25 mg. L-1 ＋KA 100μmol- L-1） group. The embryonic rat midbrain neurons were cultured for 10 days respectively. Neurons was indentificated by NSE.Subsequently, the contents of malondialdehyde（MDA） and nitric oxide （NO） in the cultured cells were measured by fluorometric assay and nitric ac- id reduction methods. Superoxide dismutase （SOD） activities were detected by xanthiuoxidase method. RESULTS：Cell growth and differentiation in GB high-dose, medium-dose and low-dose groups were significantly better than in KA group. Compared with normal control group, the activities of SOD in KA group was weakened significantly, while the content of MDA, NO were increased significantly. Compared with KA group, the activities of SOD in GB high-dose, medium-dose and low-dose groups was enhanced significantly, while the content of MDA, NO were decreased significantly.CONCLUSION： GBE can promote the growth and differentiation of embryonic rat midbrain neuronic cells and prevent the nerves from damage made by KA, which may be related with relief the lipid peroxidation in the cultured embryonic rat midbrain neurons.