大鼠创伤性脑损伤ZO-1动态变化探究.docVIP

大鼠创伤性脑损伤ZO-1动态变化探究 　【摘要】目的 探讨紧密连接蛋白ZO-1在创伤性脑损伤中的变化及意义。 方法 大鼠随机分为对照组和脑损伤组(0.5h、1h、3h、6h、24h)，参照Feeneys的自由落体致伤法，制作大鼠顶叶局灶皮质挫裂伤模型。应用伊文氏兰(Evans Blue)法测定伤后不同时间点损伤灶皮层脑组织中EB含量；应用Western blot法检测损伤灶脑微血管内皮细胞紧密连接相关蛋白ZO-1的表达水平。 结果 损伤灶皮层中EB含量在伤后1-6h明显增多，高峰期在3h。Western blot结果显示损伤后1-6h，ZO- 1表达水平较对照组显著降低（plt;0.05），3h最明显，24h与对照组无显著性差异。 结论 脑缺氧缺血时发生脑水肿，可能与ZO-1的表达减少有关。 【关键词】 创伤性脑损伤 血脑屏障 ZO- 1 【Abstract】Objective To explore the changes of tight junction proteins ZO-1 after Traumatic Brain Injury Methods Rats were randomly divided into Control group and Brain-injury group(0.5h、1h、3h、6h、24h) Brain injury animal modes were established according to Feeney’s method. Evans Blue (EB) measurement in cortex was performed to determine the content of EB in damaged cortex at different time after injury, and the protein expression of ZO-1 in microvessel endothelial cells was determined by Western blot. Results The content of EB in damaged cortex was increased obviously at the time point of 1 to 6h after injury. The extravasation peak course was at 3h after injury. Western blot revealed that the expression of tight junction (TJ) associated with protein ZO-1 was significantly decreased at the time point of 1 to 6h after injury. (Plt;0.05). and 3h was prominent. Compared with Control group, there was no significant difference in the 24h group. 毕业论文 Conclusion The decreased expression of ZO-1 may play an important role in the formation of brain edema after Traumatic Brain Injury. 【Key words】 Traumatic brain injury Blood-brain barrier ZO-1 随着国民经济和交通运输的发展，创伤性脑损伤(Traumatic Brain Injury, TBI)发病率逐年升高，其早期病理改变之一为血脑屏障(blood brain barrier, BBB)通透性增高引起的血管源性脑水肿。紧密连接（tight junction, TJ）是维持BBB完整性的重要因素，研究表明，紧密连接蛋白-1（zonula occluden-1, ZO-1）是第一个被确定的紧密连接相关蛋白，它是连接跨膜蛋白（如occludin 和claudin-5）和细胞骨架蛋白actin的桥梁[1]，是构成TJ的重要组成部分。目前，关于ZO-1在创伤性脑损伤后的动态表达研究甚少，本文旨在探讨大鼠创伤性脑损伤后不同时间点血脑屏障通透性及ZO-1表达水平的变化，以期进一步阐明脑外伤导致的BBB通透性增加的分子机制，同时为临床TBI的治疗提供新的思路。 1. 材料与方法 1.1材料与试剂 EB粉末(AMRESCO, 美国)，二甲基甲酰胺(AMRESCO, 美国)，、鼠抗ZO-1抗体（Chemicon公司）、小鼠抗β-actin单