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：《中国全科医学》2009年第4期285-287,共3页Chinese General Practice

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 ceils were randomized into groups of control （groupl）, MPP^＋ （group2）, MPP^＋ ＋EGB761 （group3）, MPP^＋ ＋LPS （group4）, andMPP^＋ ＋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 ceil 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 MTr 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^＋- in- jured 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：R651.15[医药卫生—外科学]