乳胞素诱导食蟹猴慢性帕金森模型的初步建立

Preliminary establishment of a cynomolgus monkey model of chronic Parkinson’s disease induced by lactacystin

摘要:

目的 探索通过脑室内注射蛋白酶体抑制剂乳胞素的方法建立帕金森病(PD)食蟹猴动物模型。方法 选取老龄健康雄性食蟹猴3只，随机分为实验猴2只（A猴、B猴）、对照猴1只（C猴）。实验猴多次重复脑室内注射乳胞素，对照猴注射同等剂量的0.9%氯化钠溶液。于给药前后60 min观察其行为学变化。采用免疫组织化学方法染色和行为学评分对模型进行验证。结果 行为学观察结果示，实验猴出现类似PD典型的手臂震颤、运动迟缓、运动量减少等行为学改变；行为学评分与对照猴相比，差异有统计学意义(P<0.05)。免疫组化结果示，实验猴酪氨酸羟化酶(TH)阳性面积较对照猴减少，差异有统计学意义(P<0.05)；实验猴平均吸光度较对照猴减少，差异有统计学意义(P<0.05)。实验猴α-突触核蛋白(α-syn)阳性面积较对照猴增加，差异有统计学意义(P<0.05)；实验猴平均吸光度较对照猴增加，差异有统计学意义(P<0.05)。结论 该实验采用小剂量、多次重复脑室内注射蛋白酶体抑制剂乳胞素方式产生了类似PD的行为学改变，免疫组化结果显示黑质存在PD样病变，可认为已建立食蟹猴慢性PD模型，该模型能够较好地模拟PD患者的临床症状和病理进程。 [引用格式:国际神经病学神经外科学杂志, 2021, 48(4): 321-326.]

Abstract:

Objective To establish a cynomolgus monkey model of Parkinson’s disease （PD） through intracerebroventricular injection of the proteasome inhibitor lactacystin.Methods Three elderly healthy male cynomolgus monkeys were randomly divided into experimental group， control group， and blank control group， with one cynomolgus monkey in each group. The experimental monkey was given repeated intracerebroventricular injection of lactacystin， the control monkey was given the injection of an equal dose of 0.9% sodium chloride solution， and the blank control monkey was not given any treatment. Behavioral changes were observed at 60 minutes before and after administration. Immunohistochemical staining and behavioral score were used to validate the model.Results Behavioral observation showed that the experimental monkey had the typical behavioral changes of PD such as arm tremor， bradykinesia， and reduced amount of exercise， and there was a significant difference in behavioral score between the experimental monkey and the control monkey （P<0.05）. Immunohistochemistry showed that compared with the blank control monkey， the experimental monkey and the control monkey had a significant reduction in TH-positive area， and compared with the control monkey， the experimental monkey had significant reductions in TH-positive area （P<0.05） and mean absorbance （P<0.05）. Compared with the blank control monkey， the experimental monkey and the control monkey had a significant increase in α-synuclein （α-syn）-positive area， and the α-syn-positive area in the experimental monkey was 214.5% that in the control monkey （P<0.05）； the experimental monkey had a significant increase in mean absorbance compared with the control monkey （P<0.05）.Conclusions In this experiment， repeated small-dose intraventricular injection of the proteasome inhibitor lactacystin produced the behavioral changes similar to those in PD， and immunohistochemical results showed PD-like lesions in the substantia nigra； therefore， a cynomolgus monkey model of chronic PD can be considered successfully established. This model can better simulate the clinical symptoms and pathological process of PD patients and is thus a reliable experimental animal model for studying the etiology， pathogenesis， drug therapy， and gene therapy for PD. ［Citation:Journal of International Neurology and Neurosurgery, 2021, 48(4): 321-326.］