纹状体内注射6-羟基多巴胺制备兔帕金森病模型

Making Parkinsonian model of rabbit by injecting 6-OHDA into corpus striatum

应用脑立体定位技术微量注射6-OHDA于兔右侧纹状体内。术后每周观察以阿扑吗啡(Apomorphin,APO)诱导的旋转行为,并于术后6周处死兔,以黑质酪氨酸羟化酶(Tyrosine hydroxylase,TH)免疫组化染色,观察黑质多巴胺能神经元的形态、结构及数量变化。结果表明,部分兔在术后即出现行动迟缓、躬身、易激怒等异常行为。术后6周时,20只兔中有16只在阿扑吗啡诱导后30min内的平均旋转圈数大于7r/min,达到成功模型标准。模型成功率达到80%。TH免疫组化染色可见正常对照组、假手术组及模型组未损侧黑质内有胞浆浓染、突起明显的TH免疫反应阳性神经元分布,神经元数量较多,轴突长度较长,且3者差异不显著(P>0.05);而模型组损毁侧黑质内TH免疫反应阳性神经元与上述3者相比,数目明显减少(P<0.05),残存的细胞染色较浅,胞体轮廓和突起均不清晰,轴突长度明显变短(P<0.05)。结果提示,将6-OHDA注射于兔单侧纹状体是一种制备帕金森病(Parkinson’s disease,PD)模型的有效方法,此法操作简便,动物死亡率低,模型制作成功率高。

The aim of this research was to study the method of modeling Parkinson disease（PD） by injecting 6-hydroxydopamine（6-OHDA） into right side of rabbit corpus striatum.After the surgery,the change of behavior induced by apomorphine was observed every week.The rabbits were sacrificed after six weeks and the change of morphology,structure,amount of TH positive neurons were observed in substantia nigra part using SABC immunohistochemical technique.The results showed that the abnormal behaviors,including moving stiffly,seeking for food,appeared in a part of the rabbits after operation.The rate of the apomorphin-induced rotation was over 7 r/min among 16 rabbits（80%） after 6 weeks of the injection.The rabbit with more than 7 rotation per minute was regarded as a successful PD model.TH immunohistochemical staining showed that the TH positive neurons were seen within the nigral in normal control group,sham surgery group,and the unlesioned side of model group.They had the hyperchromic cytoplasm,the clear tubers,numerous positive cells and the long axons,which had no noble diversity among above three groups（P〉0.05）.While in the opposite area of the model group TH positive neuron staining became lighter,cell body and tuber got blurrier,especially,the number and the axons length of the TH-positive neurons decreased and got shorter dramatically than that in the unlesioned side of the model group and the other two groups（P〈0.05）.The results suggest that injecting 6-OHDA into the unilateral striatum was a feasible method for establishing rabbit model of PD,which is simple manipulation with satisfying achievement ratio and lower mortality.