高压氧预适应对体外血脑屏障内皮细胞紧密连接的影响  

Effects of hypebaric oxygen preconditioning on close connection of endothelial cells in in-vitro blood-brain barrier

作　　者：郝磊[1] 邹璨 周虎传[1] 田洪[1] 张玉波[1] 宋川[1] 刘磊[1] 

机构地区：[1]解放军第三二四医院脑血管病中心,重庆400020

出　　处：《中华航海医学与高气压医学杂志》2015年第5期374-377,381,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：成都军区应用基础研究面上项目（C12024）,重庆市自然科学基金（CSTC2014jcyA 10077）

摘　　要：目的 探讨高压氧预适应对体外血脑屏障内皮细胞紧密连接的影响及机制.方法 通过hCMEC/D3细胞培养建立体外血脑屏障模型.将培养的hCMEC/D3细胞按数字表法随机分为3组：空白对照组（A组）、缺氧组（B组）及高压氧预处理组（C组）.采用Millicell-ERS系统测定血脑屏障的跨内皮阻抗（TEER）.采用细胞免疫荧光检测内皮细胞紧密连接蛋白oOccludin（≥）及ZO-1的表达变化.结果 正常培养的hCMEC/D3细胞形态呈梭形,细胞之间紧密结合,TEER值达80 ～120 Q·m^2,提示成功建立体外血脑屏障.相对于A组[（107.17±10.41）Q·m^2],B、C两组TEER值[（50.02±6.87）Ω·m^2、（83.81±8.22）Ω·m^2]均有所下降,但B组下降更明显.免疫荧光结果显示：相对于A组,B组Occludin的表达水平明显降低,而C组无明显变化.相对于A组,B组ZO-1在细胞膜表达明显降低,而胞浆表达增多,而C组ZO-1的表达无明显变化.结论 高压氧预适应可以缓解缺氧时紧密连接蛋白Occludin和ZO-1表达水平及方式的改变.Objective To explore the effects of hyperbaric oxygen preconditioning （HBOP） on close connection of endothelial cells in in-vitro blood-brain barrier and possible mechanism involved.Methods The blood-brain barrier model was developed by using hCMEC/D3 cell line.Then, the cultured cells were randomly divided into 3 groups： the blank control group （Group A）, the hypoxia group （Group B） and the HBOP group （Group C）.Millicel1-ERS was used to test TEER, while immunofluorescence was performed to detect changes in the expressions of occludin and ZO-1.Results The normal cultured hCMEC/D3 displayed a spindle shape,with the cells closely connected and the value of TEER as high as 80-120 Ω ·m^2 , indicating that the in vitro blood-brain barrier model was successfully established.Relative to the TEER value of group A [107.17 ±l0.41）Ω·m^2], theTEER values of group B and group C [（50.02±6.87）Ω ·m^2, （83.81 ±8.22） Ω ·m^2] all decreased quite considerably, with the TEER value of group B decreasing more significantly.Immunofluorenscence detection results indicated that as compared with group A, the expression level of occludin in group B decreased significantly, while no significant expression level of occludin was noticed in group C.As compared with group A, ZO-1 expression level in the cell membrane of group B decreased significantly, while the expression level of cytoplasm increased, and there were no significant changes in ZO-1 expression level in group C.Conclusions HBO preconditioning could decrease changes in the expression levels of occludin and ZO-1 induced by hypoxia.

关 键 词：高压氧预适应 血脑屏障 紧密连接 

分 类 号：R742[医药卫生—神经病学与精神病学] R459.6[医药卫生—临床医学]