高压氧通过p38丝裂原激活蛋白激酶信号通路调节大鼠脑claudin-3蛋白的表达及血脑屏障的通透性  

Effects of hyperbaric oxygen on claudin-3 in the brain tissues and blood brain barrier after cerebral ischemia-reperfusion in rats through p38 MAPK signal pathway

作　　者：张倩茹[1] 赵红[1] 卢晓梅[2] 张海鹏[2] 陈学新[3] 

机构地区：[1]中国医科大学基础医学院机能实验中心,沈阳110001 [2]中国医科大学基础医学院病理生理教研室,沈阳110001 [3]中国医科大学附属第一临床医院高压氧科

出　　处：《中华航海医学与高气压医学杂志》2011年第5期292-295,共4页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：辽宁省科技厅博士启动基金资助项目（20101149）;辽宁省教育厅高等学校科学研究项目（2005IA56）

摘　　要：目的探讨高压氧（hyperbaric oxygen，HBO）通过p38丝裂原激活蛋白激酶（p38MAPK）信号通路调节大鼠脑缺血再灌注紧密连接蛋白claudin-3的表达及血脑屏障的通透性。方法240只雄性Wistar大鼠，按数字表法随机分成5组：假手术（sham）组、脑缺血再灌注（ischemia repeffusion，IR）组、脑缺血再灌注＋HBO（IR＋HBO）组、脑缺血再灌注＋SB203580（IR＋SB203580）组、HBO＋脑缺血再灌注＋SB203580（IR＋HBO＋SB203580）组，每组48只。复制局灶性脑缺血再灌注模型，IR＋SB203580组和IR＋HBO＋SB203580组于脑缺血再灌注前30min经侧脑室注射100μl p38 MAPK信号传导通路抑制剂SB203580，IR＋HBO组与IR＋HBO＋SB203580组于再灌注期间行0．25MPaHBO治疗5次，在处死动物前1h经尾静脉注射2％伊文思蓝（Evans blue，EB），采用EB法检测缺血再灌注后血脑屏障通透性的变化。分别应用免疫组织化学的方法和Western blot法观察缺血再灌注72h后脑组织中claudin-3蛋白的分布及表达水平的变化。结果与sham组相比，其他4组再灌注后72hclaudin-3蛋白表达显著降低，差异有统计学意义（P〈0．01），同时伴有脑组织EB的含量显著增高，差异有统计学意义（P〈0．01）。IR＋HBO组（平均面密度值：0．330±0．12）和IR＋SB203580组（平均面密度值0．190±0．011）与IR组（平均面密度值：0．120±0．08）比较脑组织elaudin-3蛋白表达水平明显增加，脑组织EB的含量显著降低；与IR＋HBO组相比IR＋HBO＋SB203580组（平均面密度值：0．240±0．011）脑组织中claudin-3蛋白表达明显减少；脑组织EB的含量明显增加，差异均有统计学意义（P〈0．01，P〈0．05）。IR＋HBO＋SB203580组与IR＋SB203580组比较claudin-3含量明显增加，EB的含量明显减少，差异有统计学意义（P〈0．01）。结论HBO从蛋白水平可明显增加脑缺血再灌注脑组织中紧密连接相关蛋白claudin一3的表�Objective To investigate the effects of hyperbaric oxygen （HBO） on claudin-3 in the brain tissues and blood brain barrier （BBB） after cerebral ischemia-repeffusion in rats through p38 MAPK signal pathway. Methods Male Wistar rats were randomly divided into 5 groups, i.e. the sham, the isehemia- repeffusion （IR）, the IR ＋ HBO, the IR ＋ SB203580 and the IR ＋ HBO ＋ SB203580 groups, each consisting 48 animals. Focal cerebral isehemia-repeffusion model was reproduced. For the IR ＋ SB203580 and the IR ＋HBO ＋ SB203580 groups, 100 μl p38 MAPK signal pathway inhibitor, SB203580 was injected through the lateral ventricle 30 min before cerebral ischemia-reperfusion, and for the HBO ＋ IR and the IR ＋ HBO ＋ SB203580 groups, 0.25 MPa HBO treatment was implemented 5 times during the reperfusion period. One hour before the animals were executed, 2% Evans blue （EB） was injected into tail veins to monitor changes in the permeability of BBB following reperfusion. The distribution of claudin-3 in the brain tissue 72 hours after ischemia-reperfusion and changes in the expression of claudin-3 were monitored with Immunohistochemistry, Western blot and spectrophotometer respectively. Results When compared with the sham group, the expression of claudin-3 for the IR group decreased significantly 72 h after reperfusion（ P 〈 0.01 ） and EB content in the brain tissue increased significantly （P 〈 0. 01 ）. For the HBO ＋ IR and the IR ＋ SB203580 groups, the expression of claudin-3 in the brain increased significantly （P 〈0.01 ,P 〈0.05）, when compared with that of the IR group, and contents of EB decreased significantly （P 〈 0.01 ）. The protein expressions of claudin-3 in the IR ＋ HBO ＋ SB203580 group decreased significantly as compared with that of the IR ＋ HBO group （ P 〈 O. 01 ） and contents of EB in the brain increased significantly （ P 〈 0.01 ）. Conehlsions HBO intervention could elevate the expression of claudin-3 in the ischemia-reperfusion brain tissue,

关 键 词：高压氧 血脑屏障 脑缺血再灌注 CLAUDIN-3 SB203580 

分 类 号：R285.5[医药卫生—中药学]