银杏叶提取物对损伤后中脑多巴胺能神经细胞活性的影响  

Effects of EGB761 on Primary Cultures of Embryonic Rat Mesencephalic Cells Injured by MPP^+

作　　者：朱灿宏[1] 奚淑芳[1] 王群江[2] 周冰[1] 李蓉[1] 庄志[1] 毛善奎[1] 端礼荣[3] 

机构地区：[1]江苏省镇江市第一人民医院,212002 [2]浙江省人民医院 [3]江苏大学医学院预防医学系

出　　处：《临床合理用药杂志》2011年第05X期29-31,共3页Chinese Journal of Clinical Rational Drug Use

摘　　要：目的探讨银杏叶提取物(EGB761)对1-甲基-4-苯基吡啶离子(MPP+)及脂多糖(LPS)损伤后的中脑多巴胺能神经细胞是否具有保护作用。方法采用胚胎大鼠中脑原代细胞培养法,分离培养中脑神经细胞,培养细胞随机分为空白对照组、MPP+组、MPP++EGB761组、MPP++LPS组、MPP++EGB761+LPS组;给予MPP+、EGB761和LPS进行配组干预,然后观察细胞生长状况;通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)微量比色法检测细胞活性;酪氨酸羟化酶(TH)免疫荧光法(FITC),比较各组阳性神经元的变化,了解多巴胺能神经元的表达情况;同时硝酸还原法检测培养细胞中一氧化氮(NO)的表达情况。结果空白对照组、MPP++EGB761组:细胞生长旺盛,轴突延长明显;MPP++EGB761+LPS组:细胞生长受抑制,轴突较短MPP+组、MPP++LPS组细胞数量少,仅少数细胞稍有突起,大多数细胞无轴突。MTT法测得细胞吸光度值(A值)、免疫荧光TH阳性细胞表达及培养细胞NO的表达:空白对照组、MPP++EGB761组NO含量与MPP++EGB761+LPS组、MPP+组、MPP++LPS组比较差异有统计学意义(P<0.05)。MPP++EGB761+LPS组NO含量与MPP+组、MPP++LPS组比较差异有统计学意义(P<0.05),MPP+组与MPP++LPS组比较差异亦有统计学意义(P<0.05,见表1)。结论 LPS可加重MPP+对多巴胺能神经元的损伤作用,小胶质的活化及其释放的NO有可能参与该细胞凋亡过程。EGB761对损伤后的胚胎大鼠中脑原代培养细胞具有保护作用,此作用可能通过抑制小胶质的活化完成。Objective To explore whether ginkgo biloba extract（EGB761）could partially protect embryonic rat mesencephalic dopaminergic（DAergic）neurons injured by MPP＋.Methods Embryonic rat mesencephalic primary cell culture method was used to isolate and culture the mesencephalic neurons,and the cultured cells were randomized into groups of control（group 1）,MPP＋（group 2）,MPP＋＋EGB761（group 3）,MPP＋＋LPS（group 4）,and MPP＋＋EGB761＋LPS（group 5）.The cells were intervened with MPP＋,EGB761 and LPS to observe cell growth.Cytoactive was detected by microcolorimetry of 3-（4,5-dimethyl-thiazole-2）-2,5-diphenyl-tetrazolium Bromine-salt（MTT）,changes of positive neurons and DA neuron expression by tyrosine hydroxylase（TH） immunofluorescence（FITC）,and nitric oxide expression in cultured cells by nitric acid reduction method.Results Strong growth and significantly extended axons was noted in groups 1 and 3;inhibited cell growth and relatively short axons in group 5;a small number of cells,only a few cells with small cell processes,but most without in 2 and 4.By MTT method,cell absorbance ratio（A value）,expressions of immunofluorescence TH positive cells and of cultured cell NO were detected：groups 1,3 differed significantly from groups 5,2,4 in NO level（P〈0.05）,and group 5 from group 2,4（P〈0.05）,group 2 from group 4（P〈0.05）.Conclusion LPS can increase MMP ＋-injured DA neuron,microglia activation and the NO which it released may participate in the process of the cell apoptosis.EGB 761 may play a protective role in injured primary cultured brain cells of embryo rats,which may inhibit microglia to activate.

关 键 词：银杏叶 多巴胺 脂多糖类 帕金森病 

分 类 号：R285[医药卫生—中药学]