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

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.