MPTP偏侧恒河猴帕金森病模型研究

Establish of hemiparkinsonism rhesus monkey model by unilateral internal carotid artery injection of MPTP

目的对血管介入法、手术夹闭颈外动脉行颈总动脉穿刺、单纯颈总动脉穿刺三种甲基-苯基-四氢吡啶(MPTP)制备偏侧猴帕金森病(Parkinson's disease,PD)模型的方法进行比较,探讨偏侧猴PD模型的制作技术及效果。方法健康恒河猴6只行单侧颈内动脉MPTP注射,其中2只于DSA下行血管介入法注药;2只采用夹闭颈外动脉颈总动脉注药;2只行单纯颈总动脉注药。术后采用行为学观察和评估,包括运动障碍评分、阿扑吗啡诱发旋转实验等。猴处死后两侧黑质组织切片,NISSL和酪氨酸氢化酶(tyrosine hydroxylase,TH)免疫组织化学染色。结果血管介入法和夹闭颈外动脉颈总动脉注药法均一次建模成功;单纯颈总动脉注药法的2只分别于重复注药2次和3次后成功。行为学观察可见猴的自主运动明显减少,运动障碍评分l个月后稳定在6分以上。阿扑吗啡(apomorphine,APO)试验可诱发出向健侧旋转行为。病理检查可见注药侧黑质内神经细胞缺失减少,残留神经元变性。黑质纹状体区TH阳性多巴胺神经元较正常侧减少65%以上。结论血管介入法、颈外动脉夹闭颈总动脉注药法和单纯颈总动脉注药法均能够建立满意的偏侧猴PD模型。

Objective To investigate the skills and results of establishing hemiparkinsonism rhesus monkey models by comparing three different methods of unilateral internal carotid artery injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine （MPTP）. Methods Six healthy rhesus monkeys received MPTP injection through the internal carotid artery, among them 2 through endovascular interventional method under DSA, the other 4 through common carotid artery injection with or without external carotid artery clipping. All monkeys were evaluated after surgery according to a disability rating scale. Apomorphine （APO） evoked rotation was also tested. After death of the monkey models, both sides of substantia nigra were cut into serial sections, and tyroxine hydroxylase （TH）, immunohistochemical study was performed. Results The monkeys who received endovascular injection and common carotid artery injection with external carotid artery clipping developed symptoms of Parkinson＇s disease successfully after the first injection. Injection had to be performed 3 times and 2 times among the 2 monkeys without external carotid artery clipping. Monkeys showed PD symptoms including reduction of voluntary movement, rigidity of left limbs, bradykinesia and gait abnormality. The disability scores of hemiparkinsonian monkeys were all higher than 6 at 1 months after the last injection of MPTP. APO could evoke rotation to left. NISSL and TH immunohistochemistry staining showed that DA neurons reduced by more than 65% in the lesioned side compared with the contralateral side.Conclusion All three methods can establish satisfactory MPTP monkey models of hemiparkinsonism.