摘要
目的:观察闭锁小带蛋白-1(zonula occludens-1,ZO-1)在脑外伤后皮质中不同时段的表达变化。方法建立小鼠脑外伤模型,分为脑外伤后1h、3h、6h、12h、24h、3d、7d组,同时设立假手术组及正常对照组。脑皮质中伊文思蓝(Evans blue,EB)含量检测评估血脑屏障通透性,Western印迹法和免疫组织化学染色法检测损伤皮质区ZO-1的表达。结果脑外伤后1 h皮质中EB含量开始增加,伤后1-3 d达高峰,伤后7 d接近正常。 Western印迹法显示,ZO-1在损伤1 h后表达下调,损伤1-3 d达到最低值,损伤7 d后明显回升,但仍低于假手术组和正常对照组。免疫组织化学染色显示,正常脑皮质血管中ZO-1呈强阳性表达,损伤后表达逐渐减弱,损伤3 d后阳性表达几乎消失,之后逐渐恢复。结论 ZO-1在脑外伤后皮质区呈先降低后升高的表达规律,与脑外伤后血脑屏障通透性变化规律呈负相关,为推断脑外伤损伤时间提供了新指标。
Objective To observe the time-course expression of zonula occludens-1 (ZO-1) in cerebral cortex after traumatic brain injury (TBI). Methods The TBI model of mouse was established. The mice were divided in 1 h, 3 h, 6 h, 12 h, 24 h, 3 d, 7 d after TBI, shamand control groups. The permeability of the blood brain barrier was evaluated by measuring the extravasation of Evans blue (EB) dye. The expression of Z O-1 in cerebral cortex in the injured area was detected by western blotting and im-munohistochemistry. Results The extravasation of EBdye of injured cortex gradually increased from 1 h, peaked at 1-3 d and approximately decreased to normal at 7 d after TBI. western blotting revealed that the expression of Z O-1 gradually decreased after 1 h, was at the lowest at 1-3 d, and then significantly increased after 7 d but was still lower than that of normal and shamgroups. The result of immunohisto-chemistry showed that Z O-1 had strong expression in vessel of normal cortex, gradually decreased after TBI, and almost disappeared at 3 d after TBI and gradually recovered to normal level later. Conclusion The expression of Z O-1 in the injured cortex after TBI initially decreases and then increases. The nega-tive correlation between Z O-1 expression and EBextravasation after TBI could be used as a newindi-cator for wound age estimation.
出处
《法医学杂志》
CAS
CSCD
2015年第2期85-87,92,共4页
Journal of Forensic Medicine
基金
"十二五"国家科技支撑计划项目(2012BAK16B02)
国家自然科学基金资助项目(81273338
81172911)
上海市法医学重点实验室资助项目(14DZ2270800)
上海市博士后科研资助计划项目(14R21423000)
苏州市科技发展计划项目(SZP201304)