中文摘要：

目的：建立体外血脑屏障模型（blood-brain barrier model,BBBM）,观察非小细胞肺癌（non-small cell lung cancer,NSCLC） A549细胞对BBBM功能的影响.方法：早上将周细胞（pericytes,PC）接种到Transwell膜的外侧面,星形胶质细胞（astrocytes,AS）接种到12孔板内,8h后将Transwell置入12孔板内,再将小鼠脑微血管内皮细胞（brain microvascular endothelial cells,BMEC）接种到transwell膜的内侧面,建立血脑屏障模型.模型建立后每天进行试漏试验及跨内皮电阻（transendothelial electrical resistance,TEER）测定,鉴定是否成模.成模后分为对照组与实验组,将A549细胞接种到实验组BBBM内室,共培养体系置于培养箱,于24 h、48 h、72 h后进行电阻测定及辣根过氧化物酶渗透试验,以评估BBBM的功能.分别于渗透试验进行至2h、6h时取样,根据样本中辣根过氧化物酶浓度计算表观渗透系数（apparent permeability coefficient,Papp）.结果：（1）部分血脑屏障模型在建立后3d成模,4d可全部成模.（2） A549细胞加入BBBM内室共培养.（a）实验组TEER值与对照组比较：1d后无显著变化（P＞0.05）;2 d后降低出现统计学差异（P＜0.05）;3d后降低具有统计学差异（P＜0.01）;（b）渗透试验与对照组比较：1d后2h及6h渗透试验实验组Papp均降低,但无统计学差异（P ＞0.05）;2 d后实验组Papp升高,2h渗透试验差异具有统计学意义（P＜0.05）,6h渗透试验差异无统计学意义（P＞0.05）;3d后实验组Papp升高,2h及6h渗透试验均具有显著统计学差异（P＜0.01）.结论：小鼠体外血脑屏障模型的功能可被人类非小细胞肺癌细胞影响.体外共培养体系可用于研究肺癌脑转移过程中肿瘤细胞与脑微血管内皮细胞间相互作用的分子机制.

英文摘要：

Objective：To establish blood-brain barrier model （BBBM） in vitro,and investigate the effect of non-small cell lung cancer （A549） cells on the function of BBBM.Methods：In the morning,the pericytes were seeded on the abluminal side of Transwell membrane,and the astrocytes were seeded in 12-well plate.8 hours later the transwells were put into the wells,and the brain microvascular endothelia cells were planted on the luminal side of the transwell membrane,so that the blood-brain barrier models were established.Then the leakage test and transendothelial electrical resistance （TEER） test were performed to identify if the models are established successfully.The successful models were assigned to control group and experimental group,and the A549 cells were planted into the luminal side chamber of the experimental group BBBMs.The coculture systems were incubated in the incubator,at 24 h,48 h,72 h later,the TEER test and horseradish peroxidase permeability test were wsed to evaluate the function of BBBM.The samples of permeability test were collected when the test progress to 2 hours and 6 hours.According to the concentration of horseradish peroxidase in the samples,the apparent permeability coefficient （Papp） was calculated.Results：（1）When the constructions were performed,some of them formed BBBM successfully in 3 days,all models are formed in 4 days.（2） After the A549 cells were planted into the luminal side chamber of the BBBMs,（a） Compare to the control group：one day later,TEER of the experimental group have no significant changes （P 〉 0.05）;two days later,TEER of the experimental group reduced with statistic difference （P 〈 0.05）;three days later,TEER of the experimental group reduced with significant statistic difference （P 〈 0.01）.（b） Compare to the control group：one day later,Papp of the experimental group tend to reduce both of 2 hours and 6 hours test,but there is no statistic difference （P 〉0.05）;two days later,Papp of the experimental group incr