改良纹状体内两点注射6-OHDA法制备大鼠帕金森病模型

Improvement of rat model of Parkinson's disease by unilateral striatal two-point and two-spot injection with 6-hydroxydopamine

目的:探讨改良的单侧纹状体两点法双靶点注射神经毒素6-羟多巴胺(6-OHDA)制备帕金森病(PD)大鼠模型,对提高制模成功率的可行性。方法:利用立体定向技术以两点法双靶点注射6-羟多巴胺(6-OHDA)于雄性Sprague-Dawley大鼠右侧纹状体,分别于注射后2周和4周检测两组大鼠行为学变化;免疫组织化学法观察大鼠黑质酪氨酸羟化酶(TH)阳性细胞的数量变化;RT-PCR和Western Blot法分别检测TH mRNA和TH蛋白的表达变化。结果:造模2周后模型成功率为82.9%,4周后模型成功率高达88.6%;与对照组比较,模型大鼠6-OHDA注射侧黑质TH阳性细胞数显著减少(P<0.01);TH mRNA和TH蛋白的表达水平均明显降低(P<0.01)。结论:改良的单侧纹状体两点法双靶点注射6-OHDA制备大鼠帕金森病模型简单、易行,成功率高。

Objective： To improve the rat model of Parkinson’s disease（ PD） established by unilateral striatum twopoint and two-spot injection with 6-hydroxydopamine. Methods： Male Sprague-Dawley（ SD） rat models were established by stereotactic technique with two-point injection of 6-hydroxydopamine（ 6-OHDA） in two spots of the right striatum. After 2 and 4 weeks,the behavioral change of the rats was induced by apomorphine interperitoneal injection. The numbers of tyrosine hydroxylase（ TH） positive cells of the rat substantia nigra in the two groups were observed using immunohistochemical method. The expression of TH mRNA was examined by RT-PCR and the expression of TH protein was detected by Western Blot. Results： After 2 weeks injection of 6-OHDA in the experimental group,the successful rate of the PD model was 82. 9%. After 4 weeks injection of 6-OHDA,the success rate was up to 88. 6%. Compared with the control group,TH-positive cells were significantly decreased in rat substantia nigra of the 6-OHDA injected side（ P 〈 0. 01）,and both the expressions of TH mRNA and protein decreased significantly in the model group（ P 〈 0. 01）. Conclusion：The improved way of unilateral striatal two-point and two-spot injection of 6-OHDA to prepare the rat PD model is simple,easy and high success rate.