重组人促红细胞生成素对大鼠创伤性脑损伤后早期脑微血管内皮细胞紧密连接的保护作用。
Protective effect of rhEPO on tight junctions of cerebral microvascular endothelial cells early following traumatic brain injury in rats.
作者信息
Zhiyuan Qian, Qingyong Li, Shengming Huang, Hui Ma
机构信息
a Department of Neurosurgery , Second Affiliated Hospital of Soochow University , Suzhou , PR China.
b The Central Hospital of Xianning , Xianning , PR China.
出版信息
Brain Inj. 2016;30(4):462-7. doi: 10.3109/02699052.2015.1080386. Epub 2016 Mar 2.
OBJECTIVE
The goal of this study was to investigate the protective effect of recombinant human EPO(rhEPO) on cerebral microvascular endothelial cells and the mechanisms by which rhEPO interacts with TJs proteins, claudin-5, Occludin and ZO-1 during the early period following traumatic brain injury.
RESEARCH DESIGN
Rats (n = 81) were randomly divided into sham-operated group, TBI group and rhEPO+TBI group. Traumatic brain injury was induced by the Marmarou method.
METHODS AND PROCEDURES
Rats were killed at 3, 24, 72 and 168 hours after TBI. The integrity of the blood-brain barrier was investigated by using a spectrophotometer to assess extravasation of Evans blue dye. The expression of Claudin-5, Occludin and ZO-1 were determined by immunohistochemistry and real-time fluorescence quantitative PCR.
RESULTS
From 3 hours to 3 days, rats in the TBI group demonstrated a remarkable increase in Evans blue content in the brain, relative to rats in the sham-operated group (p < 0.05). The expression of Claudin-5 and Occludin was significantly lower than those in the sham-operated group (p < 0.05). In contrast, rats in the TBI+rhEPO group demonstrated a significant decrease in brain levels.
CONCLUSION
It was found that administration of rhEPO protected cerebral microvascular endothelial cells and reduced permeability of BBB and the mechanisms may be due to increasing the expression of TJs proteins.
目的
本研究旨在探讨重组人促红细胞生成素(rhEPO)对脑微血管内皮细胞的保护作用,以及rhEPO在创伤性脑损伤早期与紧密连接蛋白(TJs)、闭合蛋白-5(Claudin-5)、闭锁蛋白(Occludin)和紧密连接蛋白1(ZO-1)相互作用的机制。
研究设计
将81只大鼠随机分为假手术组、创伤性脑损伤(TBI)组和rhEPO+TBI组。采用Marmarou法诱导创伤性脑损伤。
方法与步骤
在创伤性脑损伤后3、24、72和168小时处死大鼠。使用分光光度计评估伊文思蓝染料的外渗情况,以研究血脑屏障的完整性。通过免疫组织化学和实时荧光定量PCR测定Claudin-5、Occludin和ZO-1的表达。
结果
从3小时到3天,与假手术组大鼠相比,TBI组大鼠脑内伊文思蓝含量显著增加(p<0.05)。Claudin-5和Occludin的表达明显低于假手术组(p<0.05)。相比之下,TBI+rhEPO组大鼠脑内水平显著降低。
结论
发现给予rhEPO可保护脑微血管内皮细胞并降低血脑屏障的通透性,其机制可能是由于增加了紧密连接蛋白的表达。