摘要
目的探讨高压氧（HBO）对局灶性脑缺血再灌注大鼠脑组织中紧密连接蛋白occludin、ZO-1蛋白表达及血脑屏障通透性的影响。方法雄性Wistar大鼠，随机分成假手术组、脑缺血再灌注组、HBO＋脑缺血再灌注组、HBO组。复制局灶性脑缺血再灌注模型，并于再灌注期间行0．25MPaHBO治疗5次，在处死动物前1h经尾静脉注射2％伊文思蓝（Evansblue，EB），采用EB法检测缺血再灌注后血脑屏障通透性的变化。应用免疫组织化学的方法观察缺血再灌注后脑组织中occludin蛋白的分布，再采用WesternBlot法检测再灌注72h后脑组织中occludin、ZO-1蛋白表达水平的变化。结果脑缺血再灌注组较假手术组脑组织EB含量显著增高（P〈0．01），其中EB的渗出以4h最为明显（1．14±0．07）U／mg。HBO＋脑缺血再灌注组较脑缺血再灌注组脑组织中EB含量显著降低（P〈0．01），HBO组与假手术组相比脑组织中EB含量变化不明显（P〉0．05）。Occludin蛋白的表达0h时相比于再灌注后4、12、24、48、72h时显著降低（P〈0．01）。现灌注72h时，HBO＋脑缺血再灌注组较脑缺血再灌注组脑组织中occludin、ZO-1蛋白表达水平于再灌注72h明显增加（P〈0．01）；HBO组与假手术组相比脑组织中occludin、ZO-1蛋白表达变化不显著（P〉0．05）。结论HBO可明显增加脑缺血再灌注脑组织中紧密连接相关蛋白occludin、ZO-1的表达，从而降低血脑屏障通透性。
Objective To investigate effects of hyperbaric oxygen （HBO） on the occludin and ZO-1 in the brain tissues and permeability of blood brain barrier （BBB） after cerebral ischemia-reperfusion in mice. Methods Male Wistar mice were randomly divided into 4 groups： the sham group, the ischemia-reperfusion group （IR）, the HBO group and the IR ＋ HBO group. Following development of the cerebral ischemia- reperfusion model, 5 sessions of HBO at 0.25 MPa were applied during the reperfusion period, and 2% Evens blue （EB） was injected into the tail veins an hour before the animals were executed. Following ischemia- reperfusion, changes in the permeability of BBB were monitored with the EB method. Following ischemia reperfusion, the distributions of occludin and ZO-1 in the brain tissue were observed with immunohistochemical method, and the expressions of occludin, ZO-1, and the contents of EB were determined by Western Blot and spectrophotometer. Results When compared with that of the sham group, the levels of EB in the brain tissue for the IR group increased significantly, reaching peak at 4 h following IR injury （P 〈 0. 01 ）. In the IR ＋ HBO group, EB contents in the brain tissue decreased significantly, when compared with those of the IR group （P 〈 0. O1 ）. However, no significant changes in the expression of occludin, ZO-1 and the content of EB in the brain tissue could be noted in the HBO, when compared with those of the sham group （P 〉 0. 05）. In the IR group, the expressions of occludin and ZO-1 at 4, 12, 24, 48, 72 h after reperfusion decreased significantly, when compared with those at 0 h（P 〈0. 01 ）. However, for the IR ＋ HBO group, the expressions of occludin and ZO- 1 at 72 h after reperfusion increased markedly（P 〈 0. 01 ）. No obvious changes in the expression of occludin and ZO-1 could be noted in the brain tissue in the animals in the HBO group, when compared with those in the sham group（P 〉 0. 05 ）. Conclusions HBO intervention could increase markedly the expressions of occludin and ZO-1 in the ischemia-reperfusion brain tissue, thus decreasing permeability of BBB.
出处
《中华航海医学与高气压医学杂志》
CAS
CSCD
2011年第2期89-92,共4页
Chinese Journal of Nautical Medicine and Hyperbaric Medicine
基金
辽宁省教育厅高等学校科学研究项目（2005IA56）
