体外MPP^+诱导胚胎大鼠中脑多巴胺能神经元模型的建立  

Establishment of the model of embryonic rat mesencephalic dopaminergic neurons induced by MPP^+

作　　者：龚超超[1] 郎娟[2] 熊中奎[1] 

机构地区：[1]绍兴文理学院医学院临床医学部,浙江绍兴312000 [2]杭州师范大学基础医学部

出　　处：《山东医药》2012年第43期17-19,共3页Shandong Medical Journal

基　　金：浙江省自然科学基金资助项目(Y2100035);浙江省公益性技术应用研究计划项目(2010C33166);浙江省教育厅高校科研计划项目(Y201018615)

摘　　要：目的建立一种体外1-甲基-4-苯基-l,2,3,6-四氢吡啶离子(MPP+)诱导的胚胎大鼠中脑多巴胺(DA)能神经元模型,为帕金森病(PD)发病机制和药物干预研究奠定基础。方法体外培养胚胎SD大鼠(孕14～15 d)中脑DA神经元7 d后随机分为实验组和对照组,均以含1%B27的无血清DMEM/F12培养基培养,在此基础上前者加入MPP+使终浓度达到10μmol/L,均继续培养48 h。采用免疫组化染色法检测酪氨酸羟化酶(TH)阳性神经元数量和突起长度。结果在放大倍数(×100)视野下,实验组和对照组中脑TH阳性神经元数量分别为(351.5±32.7)、(538.0±58.7)个,P<0.01;在放大倍数(×200)视野下,实验组和对照组中脑TH阳性神经元突起长度分别为(121.6±6.1)、(246.9±11.3)μm(P<0.001)。结论 MPP+体外诱导可成功建立胚胎大鼠中脑DA能神经元模型,此为PD发病机制和药物干预研究奠定了基础。Objective To establish a cultured cell model of embryonic rat mesencephalic dopaminergic （DA） neurons induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（ MPP＋ ）, and to lay foundation for the study of pathogenesis and drug intervention of Parkinson disease （PD）. Methods The embryonic rat （E14-15） mesencephalic dopaminergic new rons cultured in vitro for 7 days were divided into experimental group and control group, and then were both cultured with serum-free DMEM/F12 including 1% B27, the former was induced by 10 μmol/L of MPP＋ at the same time for 48 hours. The number of tyrosine hydroxylase（TH） positive neurons and neurite length were determined by immuuocytochemical ex- amination. Results Under the magnification field of 100 times ,TH （ ＋ ） neurons was 538.0 ± 58.7 in control group, and 351.5 ± 32.7 in experimental group, P 〈 O. O1 ; Under the magnification field of 200 times, the neurite length was （246.9 ± 11.3） μm in control group, and （121.6 ±6.1 ）μm in MPP＋ group（P〈0. O01）. Conclusions The model of embry- onic rat meseneephalic dopaminergie neurons can be successfully induced by MPP＋ , which could be a reliable means for latter study on pathological mechanism and drug intervention for PD.

关 键 词：多巴胺神经元 大鼠 中脑 细胞模型 1-甲基-4-苯基-l 2 3 6-四氢吡啶离子 

分 类 号：R742.5[医药卫生—神经病学与精神病学]