十溴联苯醚对血脑屏障体外模型通透性的影响  

Effect of decabromodiphenyl ether on permeability of blood-brain barrier in vitro model

作　　者：单雅楠 王舒宁 董柏茹 吴洁 SHAN Yanan;WANG Shuning;DONG Boru;WU Jie(Jinzhou Medical University,Jinzhou 121001,China)

机构地区：[1]锦州医科大学公共卫生学院,辽宁锦州121001

出　　处：《山东医药》2020年第29期32-35,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81502837);锦州医科大学大学生创新计划项目(201776)。

摘　　要：目的观察十溴联苯醚(BDE-209)对血脑屏障体外模型通透性的影响。方法利用小鼠脑微血管内皮细胞bEnd.3与星形胶质细胞构建非接触共培养血脑屏障体外模型,ERS电阻仪测定跨内皮细胞电阻值(TEER)确定建模成功,采用CCK-8法检测bEnd.3细胞活性并确定50μmol/L为BDE-209最大剂量;将0、10、25、50μmol/L的BDE-209作用于血脑屏障体外模型中的bEnd.3细胞24 h,采用ERS电阻仪测定TEER,HRP渗透实验评价模型大分子物质通透性,免疫印迹法检测bEnd.3细胞紧密连接蛋白occludin表达。结果随着BDE-209浓度增加,TEER逐渐下降(F=114.232,P<0.001),HRP渗透逐渐增多(F=103.625,P<0.001),bEnd.3细胞occludin蛋白表达量逐渐降低(F=17.291,P=0.001)。结论BDE-209可致血脑屏障体外模型TEER降低,HRP渗透增高,occludin蛋白表达减少,血脑屏障的完整性破坏。Objective To investigate effect of decabromodiphenyl ether(BDE-209)on the permeability of blood-brain barrier in vitro model.Methods In vitro model was established by unconnected co-culture of mouse cerebral microvascular endothelial bEnd.3 cells and astrocytes using Transwell chamber,and the availability was confirmed by detecting the transendothelial electrical resistance(TEER)through Millicell ERS.We used CCK-8 to detect bEnd.3 cell viability and determined 50μmol/L as the maximum dose of BDE-209.BEnd.3 cells of the in vitro model were exposed to different concentrations of BDE-209(0,10,25,and 50μmol/L)for 24 h;TEER value was measured by Millicell ERS;the permeability of macromolecules was evaluated by HRP penetration test,and the expression of tight junction protein occludin in the bEnd.3 cells was determined by Western blotting.Results With increasing BDE-209 doses,TEER decreased gradually(F=114.232,P<0.001),HRP penetration increased gradually(F=103.625,P<0.001),and occludin protein expression was down-regulated in the bEnd.3 cells(F=17.291,P=0.001).Conclusion BDE-209 could decrease TEER value,increase HRP penetration,and reduce expression of occludin protein,resulting in damage on the integrity of blood-brain barrier in vitro.

关 键 词：十溴联苯醚 血脑屏障 紧密连接 OCCLUDIN蛋白 

分 类 号：R994.6[医药卫生—毒理学]