表皮生长因子受体抑制剂AG1478联合Endophilin-1过表达增加血脑屏障通透性  

Epidermal growth factor receptor inhibitor AG1478 combined with endophilin-1 overexpression increases blood-brain barrier permeability

作　　者：刘文静[1] 王萍[2] 尚超[2] 薛一雪[2] 商秀丽[3] 

机构地区：[1]中国医科大学附属第一医院干诊神经内科 [2]中国医科大学基础医学院神经生物学教研室 [3]中国医科大学附属第一医院神经内科

出　　处：《解剖科学进展》2016年第3期306-309,314,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81100243;81502181);辽宁省科技厅科学技术计划项目(2011225020);辽宁省教育厅基金(L2013296)

摘　　要：目的研究表皮生长因子受体(EGFR)抑制剂AG1478联合Endophilin-1基因过表达对血脑屏障(BBB)通透性的影响。方法建立了体外BBB模型,实验分为四组:对照组,AG1478组,endophilin-1基因过表达(p IRES2-endophilin-1)转染组;AG1478+p IRES2-endophilin-1转染组。采用跨内皮阻抗值测定和辣根过氧化物酶渗透试验评估血脑屏障通透性变化,Western blot和免疫荧光法检测脑微血管内皮细胞中紧密连接相关蛋白ZO-1和occludin的表达和分布变化。结果与单独应用AG1478或p IRES2-endophilin-1组相比,AG1478+p IRES2-endophilin-1转染组体外血脑屏障模型跨内皮电阻值显著降低,辣根过氧化物酶透过率显著增高(<0.01),脑微血管内皮细胞中紧密连接相关蛋白ZO-1和occludin的表达水平显著降低(<0.05)。结论 EGFR抑制剂AG1478联合endophilin-1基因过表达增加BBB通透性可能与开放紧密连接相关。Objective To gain an insight into the effect of EGFR inhibitor AG1478 combined with endophilin-1 gene overexpression on the blood-brain barrier（BBB） permeability. Methods After establishing the BBB model in vitro, the experiments were randomly divided into 4 groups： control, AG1478, p IRES2-endophilin-1 and AG1478＋ p IRES2-endophilin-1 groups. Transendothelial electrical resistance（TEER） of BBB and horse radish peroxidase（HRP） flux were analyzed to evaluate the changes of BBB permeability. The expression and distribution of tight junction associated proteins ZO-1 and occludin proteins were measured by western blot and double immunofluorescence staining. Results Compared with single AG1478 or p IRES2-endophilin-1 group, TEER values were decreased, and HRP flux was increased significantly（ 〈0.01）, the expression levels of tight junction proteins ZO-1 and occludin were significantly decreased（ 〈0.05） in AG1478＋ p IRES2-endophilin-1 group. Conclusion The increased blood-brain barrier permeability by EGFR inhibitor AG1478 combined with endophilin-1 gene overexpression might be related to opening tight junction.

关 键 词：表皮生长因子受体抑制剂 endophilin-1 血脑屏障 通透性 紧密连接 

分 类 号：R741.02[医药卫生—神经病学与精神病学]