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目的探讨缝隙连接(GJ)在脑缺血再灌注血脑屏障通透性变化中的作用及其可能机制。方法①应用激光共聚焦显微镜技术观察 GJ 蛋白 Cx43在缺血再灌注半暗带区脑毛细血管周终足上含量及分布情况的变化。②将 Wistar 大鼠随机分成假手术组,手术组。线栓法制备大鼠大脑中动脉缺血再灌注模型。通过荧光分光光度定量的方法测定不同时间点脑组织中的伊文蓝含量来观察血脑屏障的通透性的改变。③取伊文蓝漏出最多的时间点,增设辛醇干预组和 DMSO 溶剂对照组,与相同时间点手术对照组比较,观察辛醇对血脑屏障通透性的影响。结果缺血2 h 再灌注3 h 脑组织伊文蓝含量开始增加,再灌注24 h 达高峰。激光共聚焦显微镜发现 GJ 蛋白 Cx43在脑内毛细血管周围的星形胶质细胞终足上分布密集。在缺血2 h 再灌注24 h 半暗带内,终足上的 Cx43分布发生变化,聚集成较大斑块。在缺血2 h 再灌注24 h 组术前给予辛醇干预,脑组织伊文蓝含量[(4.924±0.296)μg/g]低于手术对照组[(5.543±0.506)μg/g],二者差异有统计学意义(P<0.05)。结论GJ 在脑缺血再灌注后半暗带终足上分布变化明显,可能加重了血脑屏障通透性的增加;辛醇阻断 GJ可以降低脑缺血再灌注血脑屏障的通透性,从而起到减轻脑水肿的作用。
Objective To investigate the role of gap junction (GJ) in the permeability changes of blood-brain barrier after cerebral ischemia reperfusion and its possible mechanism. Methods ① Confocal laser scanning microscopy was used to observe the content and distribution of GJ protein Cx43 in the penumbra of cerebral ischemia-reperfusion area. Wistar rats were randomly divided into sham operation group and operation group. Preparation of Rat Middle Cerebral Artery Ischemia Reperfusion Model by Thread Plug Method. Fluorescence spectrophotometry was used to determine the level of Evans blue in brain tissue at different time points to observe the changes of permeability of the blood-brain barrier. ③ Take Evans Blue leaked the most time points, adding octanol intervention group and DMSO solvent control group, compared with the same time point surgery control group to observe the octanol permeability of the blood-brain barrier. Results The content of Evans blue in brain tissue began to increase at 2 h after reperfusion for 2 h and peaked at 24 h after reperfusion. Confocal laser scanning microscopy revealed that the GJ protein Cx43 was densely distributed on the final astrocytes around the capillaries in the brain. Within 24 h of reperfusion for 2 h after ischemia, the Cx43 distribution on the terminal foot changed and aggregated into larger patches. The occipitaldehyde was pretreated preoperatively at 24 h after reperfusion at 2 h after ischemia. The content of Evans blue ([(4.924 ± 0.296) μg / g] in brain tissue was lower than that of the control group [(5.543 ± 0.506) μg / g] The difference was statistically significant (P <0.05). Conclusion The distribution of GJ in the penumbra after cerebral ischemia-reperfusion may change significantly, which may aggravate the increase of permeability of blood-brain barrier. GJ can block the permeability of blood-brain barrier after cerebral ischemia-reperfusion , Which play a role in reducing cerebral edema.