快速制备鼻腔给予LPS诱导帕金森病动物模型及评价  

Quick establishment and evalmation of intranasal LPS-induced Parkinson′s disease mouse model

作　　者：宋国斌[1] 席国萍[1] 李艳花[1] 尉杰忠[1] 刘春云[1] 李凯军 柴智 马媛媛 肖保国[1,4] 张光先[1,5] 马存根 SONG Guo-Bin;XI Guo-Ping;LI Yan-Hua;YU Jie-Zhong;LIU Chun-Yun;LI Kai-Jun;CHAI Zhi;MA Yuan-Yuan;XIAO Bao-Guo;ZHANG Guang-Xian;MA Cun-Gen(Institute of Brain Science,Shanxi Datong University,Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases,Datong 037009,China)

机构地区：[1]山西大同大学脑科学研究所,神经炎症变性疾病基础与应用研究山西省重点实验室,大同037009 [2]山西中医药大学神经生物学研究中心,中西医结合防治神经系统疾病研究科技创新团队(2018TD-012),太原030619 [3]复旦大学上海医学院医学神经生物学国家重点实验室,上海200032 [4]复旦大学附属华山医院神经病学研究所,上海200025 [5]美国托马斯·杰弗逊大学神经学系,费城PA19107

出　　处：《中国免疫学杂志》2020年第11期1353-1357,1364,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(81501198,81272163);大同市应用基础研究计划项目(2018154);山西省回国留学人员重点科研资助项目(2014-重点7);山西中医学院“2011”培育计划项目(2011PY-1);神经炎症及变性疾病基础与应用研究山西省重点实验室(201805D111009)项目资助。

摘　　要：目的:经鼻腔途径给予脂多糖(LPS)可诱导帕金森病(PD)动物模型,但模型的建立需5个月之久。本课题旨在探讨一个快速建立PD模型的方法。方法:C57BL/6小鼠随机分为对照组和LPS模型组,每组9只。LPS模型组小鼠通过鼻腔滴入LPS(3 mg/kg),每侧鼻孔滴10μl,每3 d一次,连续7周,共计16次,对照组鼻腔滴入等体积生理盐水。7周后通过行为学实验检测小鼠运动能力,免疫荧光染色检测黑质多巴胺能神经元,Western blot检测黑质酪氨酸羟化酶(TH)、α-突触核蛋白的表达,HPLC检测黑质神经递质(多巴胺、3,4-二羟基苯乙酸、高香草酸、去甲肾上腺素、5-羟色胺、5-羟吲哚乙酸)含量。结果:与对照组相比,LPS模型组小鼠运动能力明显下降,黑质多巴胺能神经元数量明显减少,黑质TH表达明显降低、α-突触核蛋白的表达显著增加,黑质神经递质含量明显减少。结论:本研究通过增加每次LPS经鼻腔给药剂量、减少给药次数,建立了符合行为学及病理学特征的PD小鼠模型,并将制备过程减为7周,可明显缩短PD发病机制、治疗策略及治疗效果的研究周期。Objective:Intranasal administration of LPS can induce a mouse model of Parkinson′s disease(PD)in 5 month.Aim of this study is to accelerate process of PD model induction.Methods:Eighteen C57BL/6 mice were randomly divided into control group and LPS model group(9 in each group).Mice in LPS model group were administered by a nasal drip of LPS(3 mg/kg),10μl per nostril,once every three days for 7 consecutive weeks for a total of 16 times.Mice in control group were administered a nasal drip with same amount of normal saline.Seven weeks later,motor ability of mice was evaluated by behavioral experiment.Dopaminergic neurons in substantia nigra were observed by immunofluorescence staining.Expressions of tyrosine hydroxylase(TH)andα-synuclein in substantia nigra were detected by Western blot,and contents of neurotransmitters including DA,DOPAC,HVA,NE,5-HT,5-HIAA in substantia nigra were tested by HPLC.Results:LPS exhibited significantly reduced motor ability,accompanied by a reduction in number of dopaminergic neurons in substantia nigra,as well as in expression of TH andα-synuelein were remarkably incrased,and neurotransmitter levels in substantia nigra were decreased significantly compared with control group.Conclusion:By increasing dose and administration frequency of nasal LPS,PD model that consistent with haviour anel pathological cheracteristics can be effectively induced to 7 weeks,which can largely shorten research time for pathogenesis,treatment strategies and therapeutic effect of PD.

关 键 词：脂多糖 鼻腔给药 帕金森病 动物模型 

分 类 号：R742.5[医药卫生—神经病学与精神病学]