α-突触核蛋白原纤维诱导的帕金森病小鼠模型的行为表型及病理特征  

Behavioral and pathologic analysis of Parkinson's disease in α-synuclein fibril mice model

作　　者：田静 侯志东 任湘鹏 TIAN Jing;HOU Zhidong;REN Xiangpeng(Laboratory Center, the Eye Hospital of Wenzhou Medical University, Wenzhou 325027, China)

机构地区：[1]温州医科大学附属眼视光医院实验室中心,浙江温州325027

出　　处：《温州医科大学学报》2019年第3期157-161,共5页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金资助项目(LY18H090012);温州市公益性社会发展(医疗卫生)科技项目(Y20160016)

摘　　要：目的:研究A53T突变型α-突触核蛋白(A53T αS)原纤维诱导的帕金森病(PD)小鼠模型的行为表型及病理学特征。方法:小鼠脑内黑质致密区(SNpc)定位注射5 μg A53T αS建立PD小鼠模型,造模3个月后,通过旷场试验、悬挂试验和Y迷宫检测模型小鼠的运动和认知行为学表型。分别使用抗磷酸化α-突触核蛋白(p Syn)、抗酪氨酸羟化酶(TH)及小胶质细胞特异性蛋白抗体IBA-1作为评价模型小鼠脑内的路易小体包涵体、多巴胺能神经元变性死亡及神经炎症的分子标记,通过免疫组织化学染色法研究模型小鼠脑内典型的病理特征。结果:旷场试验中,模型组小鼠的总运动距离较对照组差异无统计学意义(P>0.05);悬挂试验显示模型组小鼠四爪抓杆持续时间较对照组小鼠显著缩短(P<0.05);Y迷宫检测表明模型组小鼠的自发转换正确率较对照组差异无统计学意义(P>0.05)。与对照组比,模型组小鼠中脑SNpc区的pSyn阳性包涵体数量显著增多(P<0.01),同时TH阳性神经元数量显著减少(P<0.05),IBA-1阳性胶质细胞数量异常增加(P<0.01)。结论:脑内注射A53T αS原纤维可稳定诱导出PD小鼠模型,该模型小鼠表现出一定程度的肌力和平衡力下降;且可同时模拟出PD的路易小体包涵体、多巴胺能神经元缺失及过度激活的神经炎症等病理特征。Objective: To study the behavioral phenotype and pathological features of A53T mutant α-synuclein (A53T αS) fibril mice model of Parkinson’s disease (PD). Methods: A53T αS fibril PD model was established by injecting 5 μg of A53T αS into the substantia nigra pars compacta (SNpc) of mice brain for 3 months. The motor and cognitive behavioral phenotypes of the model mice were studied by open field test, hanging test and Y maze. Anti-phosphorylation of α-synuclein (pSyn), anti-tyrosine hydroxylase (TH) and microgliaspecific protein antibody IBA-1 were used as molecular markers of Lewy bodies (LBs), dopamine (DA) neurons and neuroinflammation, respectively. These markers were used for immunohistochemical staining to evaluate the typical pathological characteristics in the brain of PD model mice. Results: In the open field test, total locomotive distance was almost equal in both groups of mice (P>0.05);however, duration of the four-jaw grip of the model mice in the hanging test was much shorter than that of the control mice (P<0.05). In the Y maze test, the correct rate of spontaneous alterations of the model mice was a bit lower than that of the control mice, but there was no significant difference (P>0.05). Compared with the control mice, the number of pSyn-positive inclusions and IBA-1 positive cells in the SNpc region of the model mice were markedly increased (P<0.01), whereas the number of TH-positive neurons was notably decreased (P<0.05). Conclusion: Intra-nigral injection of A53T αS fibrils can stably induce a mouse model of PD, which exhibits a certain decline in motor function and can simul taneously recapitulates the typical pathological features of PD, such as LBs, the degeneration of DA neurons, and over-activated neuroinflammation as well.

关 键 词：帕金森病模型 α-突触核蛋白原纤维 行为学 路易小体 多巴胺能神经元 小鼠 

分 类 号：Q953[生物学—动物学] R742.5[医药卫生—神经病学与精神病学]