提高帕金森病动物模型制作成功率的方法学探讨及模型综合评价

目的探讨提高6-OHDA帕金森病(PD)大鼠模型制作成功率的方法,并对模型进行综合评价。方法取SD大鼠90只,将6-OHDA立体定向注射于左侧黑质区及中脑腹侧被盖,观察大鼠的行为、黑质抗氧化指标、线粒体呼吸链及黑质细胞形态学的变化。结果①经阿朴吗啡诱导后共筛选出成功模型64只,成功率为71%;②模型大鼠黑质区ROS、MDA水平均明显升高,而GSH-Px活性则明显降低;③模型大鼠黑质区存在线粒体呼吸链功能障碍;④免疫组化发现注射侧黑质区多巴胺能神经元较对侧明显减少(P<0.001);⑤电镜观察发现模型大鼠黑质细胞同时存在凋亡、变性和坏死样改变。结论应用本方法可较快建立稳定的成功率较高的PD大鼠模型,该方法是通过增强氧化应激、干扰线粒体呼吸链功能、诱导凋亡甚至引发坏死等不同机制损毁多巴胺能神经元的,同PD的发病机制基本一致。

Establishment of an improved rat model of Parkinson' s disease

Objective To study the method of increasing success rate of Parkinson's disease (PD) rat model establishment. Methods Ninety SD rats were administered with stereotaxic microinjection of 6-OHDA into the left substantia nigra compact part (SNC) and the ventral tegmental area (VTA) of the midbrain. The changes of behaviors, nigral antioxidation index, mitochondrial respiration-chain function, and the morphology of nigra dopaminergic neuron (NDN) were observed after 6-OHDA injection. Results PD rat model was successfully established in 64 rats, with success rate of 71%. In the rat models, the contents of ROS and MDA obviously increased but GSH-Px activity significantly decreased. Dysfunction of mitochondrial respiration-chain was noted in substantia nigra. The immunohistochemical staining showed a significant reduction of TH-positive neurons in the 6-OHDA injection side compared with the opposite side. Electron-microscope demonstrated that apoptosis, degeneration and necrosis existed extensively in NDN. Conclusions PD rat model with high success rate was established by two site injection of 6-OHDA which could damage NDN probably through increasing oxidative stress, interfering mitochondrial respiration-chain function, and inducing apoptosis and necrosis.