Biphasic activation of nuclear factor kappa B and expression of p65 and c-Rel after traumatic brain injury in rats
Biphasic activation of nuclear factor kappa B and expression of p65 and c-Rel after traumatic brain injury in rats
复制标题
大鼠脑外伤后核因子 kappa B 的双相激活以及 p65 和 c-Rel 的表达。
DOI：
10.1007/s00011-013-0677-1
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发表时间：
2014-02-01
影响因子：
6.7
通讯作者：
Hang, Chun-Hua
中科院分区：
文献类型：
--
作者：
Hu, Yang-Chun;Sun, Qing;Hang, Chun-Hua
核因子-κ B（NF-κ B）作为中枢神经系统中的关键调节因子发挥作用并调节炎症途径。新生儿缺氧缺血和蛛网膜下腔出血后脑内NF-κ B B活化有两个高峰。我们的前期研究发现，创伤性脑损伤（TBI）后NF-κ B B活性在早期即上调，并在第7天仍保持较高水平。然而，缺乏关于TBI后NF-κ B活性和NF-κ B亚基表达的综述数据。因此，本研究旨在观察创伤后不同时间点脑挫伤区NF-κ B B激活及NF-κ B B p65和c-Rel亚单位表达的变化。建立大鼠TBI模型，采用凝胶电泳迁移率改变法检测脑损伤周围组织NF-κ B B DNA结合活性。Western blotting检测细胞核中p65和c-Rel蛋白的表达。采用酶联免疫吸附法检测肿瘤坏死因子-α（TNF-α）和白细胞介素-1 β（IL-1 β）的浓度。免疫组化检测c-Rel和p65的分布，脑皮质NF-κ B B活性在伤后3天和10天出现双峰。此外，发现p65蛋白水平升高，并在TBI后3天达到峰值，而c-Rel水平在损伤后期显著升高。实验性脑损伤后TNF-α和IL-1 β水平呈双相性升高，NF-κ B B呈双相性激活。NF-kappa B p65和c-Rel亚基在TBI后不同时间段升高，导致不同的NF-kappa B亚基可能参与TBI后不同的病理生理过程的假设。
Nuclear factor kappa B (NF-kappa B) functions as a key regulator in the central nervous system and regulates the inflammatory pathway. There are two peaks of cerebral NF-kappa B activation after neonatal hypoxia-ischemia and subarachnoid hemorrhage. Our previous studies found that NF-kappa B activity was up-regulated at an early stage and remained elevated at day 7 after traumatic brain injury (TBI). However, data are lacking regarding an overview of NF-kappa B activity and expression of NF-kappa B subunits after TBI. Hence, the current study was designed to detect the time course of NF-kappa B activation and expression of NF-kappa B p65 and c-Rel subunits around the contused cortex following TBI.Adult Sprague-Dawley rats were randomly divided into sham and TBI groups at different time points. A TBI model was induced, and then the NF-kappa B DNA-binding activity in the surrounding areas of injured brain was detected by electrophoretic mobility shift assay. Western blotting was used to measure the protein levels of p65 and c-Rel in the nucleus. The concentrations of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) were detected by enzyme-linked immunosorbent assay. Moreover, the distribution of c-Rel and p65 was examined by immunohistochemical studies.There were double peaks of cerebral cortical NF-kappa B activity, at 3 and 10 days post-injury. Additionally, protein levels of p65 were found to be elevated and peaked at 3 days after TBI, while levels of c-Rel were elevated significantly during the later phase of injury. Furthermore, TNF-alpha and IL-1 beta concentrations also showed a biphasic increase.Biphasic activation of NF-kappa B could be induced after experimental TBI in rats. NF-kappa B p65 and c-Rel subunits were elevated at different post-TBI time periods, leading to a hypothesis that different NF-kappa B subunits might be involved in different pathophysiological processes after TBI.