Objective To establish a cultured cell model of embryonic rat mesencephalic dopaminergic (DA) neurons induced by l-methyl-4-phenyl-l ,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 neurons 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 (xmol/L of MPP+ at the same time for 48 hours. The number of tyrosine hydroxylase ( TH) positive neurons and neurite length were determined by immunocytochemical examination. 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 <0. 01; Under the magnification field of 200 times, the neurite length was (246. 9 ±11.3) p,m in control group, and (121.6 ±6. 1 ) u,m in MPP+ group( P <0.001). Conclusions The model of embryonic rat mesencephalic dopaminergic neurons can be successfully induced by MPP* , which could be a reliable means for latter study on pathological mechanism and drug intervention for PD.%目的 建立一种体外1-甲基-4-苯基-1,2,3,6-四氢吡啶离子(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7d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10 μmol/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.
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