应用6-羟基多巴胺建立帕金森病大鼠模型的稳定性评价

背景应用6-羟基多巴胺(6-Hydroxydopamine,6-OHDA)损毁黑质多巴胺神经元(nigral dopaminergic neuron,NDN)制作的大鼠模型,在目前帕金森病的研究中应用最广,但由于黑质致密部体积狭小,技术难度大,故模型制备成功率一般仅为30％～40％左右.目的探讨提高6-羟基多巴胺帕金森病大鼠模型成功率的方法,并对模型进行评价.设计完全随机的实验研究.地点和材料实验在安徽中医学院第一附属医院中心实验室完成,实验材料包括SD大鼠,6-OHDA,阿朴吗啡,兔抗TH血清,ABC试剂盒,SR-6N大鼠脑立体定向仪,JEM-100CX电镜.干预取SD大鼠90只,将6-OHDA立体定向微量注射于左侧黑质区及黑质纹状体通路,观察大鼠行为及黑质细胞形态学变化.主要观察指标旋转行为,免疫组织化学观察多巴胺能神经元数量,电镜观察多巴胺能神经元形态改变.结果90只大鼠中经阿朴吗啡诱导后有64只(占71％)恒定转向右侧且结果稳定,旋转圈数30 min＞210 r,被视为成功大鼠模型;免疫组化观察发现注射侧黑质区多巴胺能神经元较对侧明显减少,电镜观察发现其普遍存在凋亡及坏死样改变.结论应用本方法可较快建立稳定的成功率较高的大鼠模型,但在病理和行为学等方面同自然患者仍有较多差异.

Reliability of a 6-OHDA rat model of Parkinson's disease

BACKGROUND: The most commonly used animal model of Parkinson's disease is created by lesion of nigral dopaminergic neuron (NDN) resulted from intrastriatial injection of 6-hydroxydopamine (6-OHDA) . However, the success rate is as low as 30% - 40% because of the tiny size of the striate.OBJECTIVES: To investigate the feasibility of a new method using 6-OHDA to create Parkinson model in rats with a higher success rate.DESIGN: Full-randomized experimental study.SETTING AND MATERIALS: The experiment was finished in the Central Laboratory, the First Affiliated Hospital of Anhui College of Traditional Chinese Medicine. SD rats, 6-OHDA, Apomorphine, rabbit anti-TH serum, ABC kit,SR-6N stereotaxic instrument and JEM-100CX electron microscope.INTERVENTION: Ninety SD rats received left side intrastriatal stereotaxic injection of 6-OHDA. Behavioral analysis of turning and morphological analyses on quantity and structure of doparminergic neurons (DN) were performed.MAIN OUTCOME MEASURES: Rotation behavior, the number of dopaminergic neurons by immunohistochemistry, and the neurological changes of dopaminergic neurons by electron microscope.RESULTS: A success Parkinson model was considered when the level of turning induced by apomorphine above 210 turns per 30 minutes. The success rate was 71% (Parkinson model in 64 out of 90 rats). Immunohistochemistry revealed decreased number of DNs in the injection side than that in the contralateral side. Electron microscopy(EM) found apoptosis and necrosis in the injection area.CONCLUSIONS: This new method is able to create reliable Parkinson's disease model in rats in a high success rate, although the pathology and behaviour of Parkinson rats differ somehow from those of natural Parkinson's disease.