高温对血脑屏障内皮细胞紧密连接的作用及其机制  

Effect of hyperthermia on tight junctions of endothelial cells in the in vitro blood-brain barrier model and its molecular mechanism

作　　者：陈一招[1] 徐如祥[1] 杨志林[1] 徐宗俊[2] 姜晓丹[1] 蔡颖谦[1] 

机构地区：[1]第一军医大学附属珠江医院神经外科,广州510282 [2]第一军医大学附属珠江医院检验科,广州510282

出　　处：《中华创伤杂志》2003年第9期540-543,共4页Chinese Journal of Trauma

基　　金：国家自然科学基金资助项目 ( 3 0 17961);军队十五重点项目基金资助项目 ( 0 1ZD5 4)

摘　　要：目的　探讨高温对血脑屏障内皮细胞间紧密连接的影响及其分子机制。　方法　利用内皮细胞与胶质细胞共培养建立体外血脑屏障模型。采用Millicell-ERS系统检测血脑屏障模型的跨内皮阻抗 (transendothelialresistance,TER)。对内皮细胞间紧密连接采用银染的方法研究其结构的完整性。运用半定量逆转录 -聚合酶链式反应 (RT -PCR)及Westernblotting的方法分析高温作用后血脑屏障内皮细胞紧密连接蛋白 (zonulaoccluden 1,ZO - 1)与occludin的表达变化。　结果　 4 3℃高温作用 2h后 ,体外血脑屏障模型TER均值由 (32 1.30± 5 8.5 9)Ω·cm2 下降至 (6 5 .6 7± 6 .0 2 )Ω·cm2 。同时 ,内皮细胞紧密连接完整性明显破坏 ,内皮细胞ZO - 1及occludin的表达水平明显降低。　结论　高温可以明显破坏血脑屏障内皮细胞间紧密连接的完整性 ,高温条件下内皮细胞ZO - 1与occludin表达水平的降低是血脑屏障热损伤的重要分子机制。Objective To discuss the effect of hyperthermia on tight junctions of the endothelial cells in the blood-brain barrier and explore the molecular mechanism. Methods An in vitro blood-brain barrier model was established by coculture of ECV304 and astrocytes. Transendothelial resistance (TER) of in vitro blood-brain barrier was determined by Millicell-ERS system. The morphological change of tight junctions of the endothelial cells in the in vitro blood-brain barrier was determined by the method of silver staining. The expression levels of zonula occluden 1 (ZO-1) and occludin were analyzed by means of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western bloting. Results After two hours at 43℃, the mean value of TER was decreased from (321.30± 58.59) Ω·cm2 to (65.67±6.02) Ω·cm2. The integrity of tight junctions was destroyed and the expressions of ZO-1 and occludin decreased significantly. Conclusions Hyperthermia can destroy the tight junctions of the endothelial cells in the in vitro blood-brain barrier. The expression decrease of ZO-1 and occludin induced by hyperthermia is one of the most important molecular mechanisms.

关 键 词：血脑屏障 内皮细胞 紧密连接 高温 紧密连接蛋白 OCCLUDIN 

分 类 号：R782.4[医药卫生—口腔医学]