帕金森病模型鼠黑质α-突触核蛋白表达升高并集聚  

Up-regulation and aggregation of α-synuclein in mouse model of parkinsonism

作　　者：杜芸兰[1] 刘振国[1] 陈生弟[1] 李彪[2] 陆国强[1] 

机构地区：[1]上海第二医科大学附属瑞金医院神经内科,神经病学研究所,200025 [2]上海第二医科大学附属瑞金医院核医学科,200025

出　　处：《中华神经科杂志》2005年第6期359-362,共4页Chinese Journal of Neurology

基　　金：上海市教委发展基金资助项目(03BK24);上海市科学发展基金项目合同(01QMH1410)

摘　　要：目的观察低剂量1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)对小鼠黑质多巴胺能神经元α-突触核蛋白表达的影响。方法C57BL小鼠腹腔注射MPTP20mg/kg体重,每周注射1次连续3周,观察小鼠行为改变,应用逆转录-聚合酶链反应(RT-PCR)、Western印迹法、免疫组化染色检测α-突触核蛋白表达,荧光双标染色观察多巴胺能神经元内α-突触核蛋白聚集。结果与注射生理盐水组相比,注射MPTP组小鼠出现暂时性躯干震颤、竖毛、动作减少等;α-突触核蛋白基因表达百分比(79·5%±0·8%,生理盐水组为19·2%±3·3%,P<0·05)及蛋白表达百分比(82·2%±4·1%,生理盐水组为18·5%±1·0%,P<0·05)均明显增加;黑质部酪氨酸羟化酶(TH)阳性细胞数减少,α-突触核蛋白阳性细胞数增加;α-突触核蛋白与硫磺素S、TH与硫磺素S复合定位染色的阳性细胞数均升高。结论低剂量MPTP可诱导小鼠行为改变,黑质多巴胺能神经元α-突触核蛋白表达升高,并出现蛋白聚集。Objective To investigate α-synuclein expression in dopaminergic neurons of mouse exposed to low dose of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Methods The C57BL mice received intraperitoneal injections of 20 mg/kg MPTP once a week for 3 consecutive weeks. Abnormal behavior was observed during the treatment by using herbicide. The expression of α-synuclein was determined with RT-polymerase chain reaction (RT-PCR),Western blot and immunohistochemistry. The aggregation of α-synuclein was assessed with double immunofluoresence staining. Results As compared with the controls of mice,those treated with MPTP for 3 weeks displayed markedly hypoactive behavior and the percentage of gene and protein expression of α-synuclein were 79.5 %±0.8% and 82.2%±4.1%,while those of the control group were 19.2%±3.3% and 18.5%±1.0%, respectively(P<0.05). Immunohistochemical staining showed that the density of α-synuclein stain was significantly increased after treatment with MPTP and localized in the TH immunoreactive neurons. The colocalization positive neurons with thioflavine S and α-synuclein, thioflavine S and TH, were observed in the double labeling immunofluorescence with confocal microscope. Conclusions MPTP might induce up-regulation and aggregation of α-synuclein in dopaminergic neurons of substantial nigra.

关 键 词：核蛋白表达 逆转录-聚合酶链反应(RT-PCR) 升高 帕金森病 黑质多巴胺能神经元 WESTERN印迹法 Α-突触核蛋白 模型鼠 生理盐水组 C57BL小鼠 集聚 蛋白基因表达 酪氨酸羟化酶 行为改变 蛋白聚集 细胞数 四氢吡啶 腹腔注射 

分 类 号：R742.5[医药卫生—神经病学与精神病学]