微流控芯片上的神经元损伤模型的建立及相关药物的保护作用  

Construction of Neuron Injury Model and Research of Relevant Neuronal Protective Drugs on Microfluidic Chips

作　　者：雷鸣 张玉洁 李银萍[3] 雷祎凤 LEI Ming;ZHANG Yujie;LI Yinping;LEI Yifeng(Dept. of Neurology, First People＇s Hospital of J iangxia District, Wuhan 430200, China;Institute of Technological Sciences, Wuhan University, Wuhan 430072, China;School of Basic Medical Sciences, Wuhan University, Wuhan 430071 , China)

机构地区：[1]武汉市江夏区第一人民医院神经内科,湖北武汉430200 [2]武汉大学工业科学研究院,湖北武汉430072 [3]武汉大学基础医学院,湖北武汉430071

出　　处：《武汉大学学报（医学版）》2018年第4期610-614,共5页Medical Journal of Wuhan University

基　　金：国家自然科学基金(编号:31500775);中央高校基本科研业务费专项基金(编号:2042018kf0235);武汉市青年科技晨光计划(编号:2017050304010306);武汉大学珞珈青年学者计划

摘　　要：目的：在微流控芯片中,构建神经元损伤模型,探讨鼠神经生长因子和奥拉西坦对受损神经元的保护作用。方法：在区室化微流控芯片中培养神经元,诱导神经突的定向生长;在芯片通道中用6-羟基多巴胺（6-OHDA）诱导神经元损伤;然后随机分为3组：试验组1给予鼠神经生长因子培养7d,试验组2给予奥拉西坦培养7d,对照组不添加药物培养7d。用光学显微镜和荧光显微镜观察不同治疗组的神经元细胞生长,用蛋白印迹检测神经元特异性微管蛋白Tuj1表达水平。结果：治疗7d后,两组试验组的神经元生长情况均明显优于对照组、神经突密度较对照组大幅提高、并且Tuj1蛋白表达水平明显升高;试验组1治疗后的神经元生长情况优于试验组2。结论：在微流控芯片上的体外神经元损伤模型中,鼠神经生长因子和奥拉西坦对受损神经元具有显著保护作用,并且鼠神经生长因子的保护作用优于奥拉西坦。该芯片将可能为神经元相关保护药物的开发和体外筛选提供新思路和新研究技术。Objective：To establish an in vitro neuronal injury model on a microfluidic chip,and to study the protective effect of mouse nerve growth factor（NGF）and oxiracetam on chip.Methods：We incubated the neurons on the microfluidic compartmentalized chips in order to induce the directional growth of neurites,6-hydroxydopamine（6-OHDA）was introduced in the chips to establish the in vitro neuronal injury model.Then we separated the injured neurons into 3 groups,the 1 st group was incubated with mouse NGF for 7 days,the 2 nd group was treated with oxiracetam for7 days,and the control group was incubated without the addition of drugs.We examined the neuronal growth under optical microscope and fluorescence microscope,then evaluated the expressionof Tuj1 protein with Western blot.Results：After incubation of 7 days,both two treatment groups enhanced the neuronal growth,promoted the neurite density,and increased the expression level of Tuj1.Moreover,the neuronal growth of the 1 st group was better than the 2 nd group.Conclusion：On the in vitro neuronal injury model on microfluidic chip,both mouse NGF and oxiracetam have significantly protective effect on injured neurons,and the protective effect of NGF may be better than oxiracetam.This microfluidic chip may provide a useful platform for investigation and screening of new drugs with neuronal protective effect.

关 键 词：微流控芯片 神经元损伤 鼠神经生长因子 奥拉西坦 神经保护 

分 类 号：R318[医药卫生—生物医学工程]