MPTP诱导慢性帕金森病恒河猴模型的初步建立  

Preliminary establishment of chronic Parkinson's disease in rhesus monkey model induced by injection of MPTP

作　　者：史良琴 罗启慧[1,2] 曾文 龚立 程安春[1,2] 毕凤君 曾利才 陈姗姗[1,2] 陈正礼[1,2] 

机构地区：[1]四川农业大学动物医学院,动物疾病与人类健康中心四川省重点实验室,四川雅安625014 [2]四川农业大学预防兽医研究所,成都611130 [3]四川普莱美生物科技有限公司/国家实验猕猴种源基地,四川雅安625014

出　　处：《浙江大学学报（农业与生命科学版）》2014年第3期257-265,共9页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：四川省科技厅应用基础项目(2011JY0054);"十二五"国家科技重大专项(2011ZX09301-001);四川省青年科技创新研究团队项目(2013TD0015)

摘　　要：拟建立一种不仅操作简便而且能较好模拟帕金森病(Parkinsons disease,PD)临床症状和病理进程的造模方法。选取健康老龄雌性恒河猴12只,随机分为实验组9只(进而按临床症状及行为学评分又分为3个亚组)和对照组3只,以0.2mg/(kg·d)小剂量、多次重复肌内注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)来建立慢性PD恒河猴动物模型,对照组注射同等剂量的0.9%氯化钠溶液。每日于MPTP给药前后观察记录各组动物行为学表现各30min。实验结束后,全部处死动物,采用免疫组织化学法检测黑质酪氨酸羟化酶(tyrosine hydroxylase,TH)和α-突触核蛋白(α-synuclein,α-syn)的分布与表达变化对模型进行验证。行为学结果显示,实验组动物出现3种不同程度的临床表现,根据其轻重,将其分为3个亚组;TH免疫组织化学结果显示,阳性物质主要分布于神经元胞质和突起内,与对照组相比,实验组中的第1、第2、第3亚组阳性总面积分别减少71.90%,61.90%,45.74%,差异有统计学意义(P<0.01);α-syn免疫组织化学结果显示,阳性物质主要分布于神经突起内,其次为细胞间质,并在濒临死亡动物的黑质致密部检测到路易小体,实验各组动物黑质阳性总面积较对照组分别增加170.29%,137.82%,47.88%,差异有统计学意义(P<0.01)。上述结果表明,采用小剂量、多次重复肌内注射MPTP方式能够建立老龄恒河猴PD动物模型,该模型能够较好地模拟PD病人的临床症状和病理进程,是研究PD病因、发病机制、药物治疗及基因治疗较可靠的实验动物模型。Summary Parkinson＇ disease（PD） is a common and age-related progressive neurodegenerative disorder,and its pathogenesis is not yet entirely clear.The present study is to establish a model easy to control and better simulate the clinical symptoms,processes and pathological changes of PD patients,expecting to provide a foundation and platform service for pathogenesis and treatment research of Parkinson＇s disease. Twelve healthy aging female rhesus monkeys,divided into experimental group （nine rhesus monkeys） and control group （three rhesus monkeys） randomly,were respectively daily injected a small dose（0.2 mg/（kg？d））of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP） and the same dose of saline by intramuscular injection for 45 days repeatedly.The behavioral manifestations of all monkeys were evaluated before and after the injection for 30 min,respectively.After sacrificed,the expression and distribution of tyroxine hydroxylase （TH） and α-synuclein （α-syn） in substantia nigra were studied by immunohistochemical method,and the immunopositive total area and average optical density were used to validate the model. The results of behavioral manifestations indicated that,experimental animals showed three different degrees of clinical manifestations and could be divided into three subgroups （three animals each subgroup） according to the behavioral and clinical manifestations.The first subgroup showed typical behavioral manifestations of Parkinson＇ disease and one animal was in the moribund period.The activity,posture and bradypraxia of the third subgroup were slightly abnormal,and behavioral manifestations of the second subgroup were between the first and the third subgroup.However the control group did not show any abnormality.The immunopositive productions of TH were mainly distributed at the cytoplasm and neurites.Compared with the control group,the positive cytoplasm and neurites of the first subgroup were severely reduced,even no positive production was observed i

关 键 词：1-甲基-4-苯基-1 2 3 6-四氢吡啶 帕金森病 黑质 酪氨酸羟化酶 Α-突触核蛋白 免疫组织化学 恒河猴 

分 类 号：S858.23[农业科学—临床兽医学] R332[农业科学—兽医学]