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目的建立大鼠帕金森病模型,观测其行为学、中脑黑质多巴胺能神经元及超微结构的变化。方法利用立体定向技术,注射6-OHDA至大鼠中脑黑质SNc和中脑腹侧被盖区VTA,于注射后2、4、6、8周观测阿朴吗啡诱导的大鼠旋转行为学变化;采用免疫组化染色检测TH的表达,了解中脑黑质多巴胺能阳性神经元变化;利用透射电镜了解黑质区超微结构的变化。结果所有大鼠进行阿朴吗啡诱导旋转行为测试,旋转圈数>7r/min,并且>210r/30min,为合格的PD大鼠模型。大鼠模型于第4、6和8周时,大鼠行为学变化较为恒定。第8周时,50只大鼠中出现32只大鼠大鼠旋转次数平均为11.5±1.2/分,造模成功率为64%。PD模型大鼠超微结构观察,黑质神经元数目明显减少,线粒体肿胀,粗面内质网囊性扩张、脱颗粒,髓鞘扩张。免疫组化检测,成功模型光镜下分别计数双侧黑质TH阳性神经元,损毁侧黑质TH阳性神经元占正常侧的3.0%,即毁损侧TH阳性神经元减少97.0%。结论利用立体定向技术,选择黑质和中脑腹侧被盖区等双靶点,可建立6-OHDA大鼠PD模型,具有明确病理变化,提高模型成功率。
OBJECTIVE: To establish a rat model of Parkinson’s disease and observe the changes of its behavior, dopaminergic neurons and ultrastructure in substantia nigra of midbrain. Methods Stereotactic technique was used to inject apomorphine-induced rat spinach behavior by injecting 6-OHDA into substantia nigra SNc and mesencephalic ventral tegmental area VTA at 2,4,6,8 weeks after injection The expression of TH was detected by immunohistochemical staining to understand the changes of dopaminergic neurons in substantia nigra of midbrain. The ultrastructure of substantia nigra was observed by transmission electron microscopy. Results All rats were subjected to apomorphine-induced rotational behavior test with> 7 r / min rotation and> 210 r / 30 min, which was a qualified PD rat model. At the 4th, 6th and 8th week of rat model, the behavioral changes in rats were relatively constant. At the 8th week, 32 rats in 50 rats appeared an average of 11.5 ± 1.2 / rotation, and the success rate of modeling was 64%. PD model rats ultrastructural observation, significantly reduced the number of neurons in the substantia nigra, mitochondria swelling, cytoplasmic expansion of rough endoplasmic reticulum, degranulation, myelin dilation. Immunohistochemical detection and successful model respectively counted bilateral substantia nigra TH positive neurons under light microscope. The number of TH substantia nigra neurons on the lesioned side accounts for 3.0% of the normal side, ie, the number of TH positive neurons on the lesioned side is reduced by 97.0%. Conclusion The stereotactic technique was used to select the target sites of substantia nigra and mesencephalic ventral tegmental area to establish the PD model of 6-OHDA rats with clear pathological changes and to improve the model success rate.