Novel effects of edaravone on human brain microvascular endothelial cells revealed by a proteomic approach.
Novel effects of edaravone on human brain microvascular endothelial cells revealed by a proteomic approach.
复制标题
通过蛋白质组学方法揭示依达拉奉对人脑微血管内皮细胞的新作用。
DOI：
10.1016/j.brainres.2013.08.019
复制
发表时间：
2013
期刊：
影响因子：
2.9
通讯作者：
Kato T.
中科院分区：
文献类型：
--
作者：
Onodera H;Arito M;Sato T;Ito H;Hashimoto T;Tanaka Y;Kurokawa MS;Okamoto K;Suematsu N;Kato T.
依达拉奉（3-甲基-1-苯基-2-吡唑啉-5-酮）是一种自由基清除剂，用于急性缺血性卒中。然而，目前尚不清楚依达拉奉是否仅作为自由基清除剂或具有其他药理作用。因此，我们采用二维荧光差异凝胶电泳（2D-DIGE）技术研究了依达拉奉对人脑微血管内皮细胞（HBMEC）的作用。共发现38个蛋白质点，依达拉奉处理后其强度平均显著改变1.3倍（p< 0.05），并成功鉴定了其中的17个蛋白质，其中4个为细胞骨架蛋白或细胞因子调节蛋白。因此，我们随后通过免疫细胞化学研究了HBMEC的大小和形状，肌动蛋白网络和紧密连接的变化。因此，与未处理的HBMEC相比，大多数经依达拉奉处理的HBMEC变得更大更圆。此外，依达拉奉处理的HBMEC沿细胞连接处沿着形成聚集性紧密连接蛋白（ZO）-1。此外，我们发现依达拉奉抑制白细胞介素（IL）-1β诱导的单核细胞趋化蛋白-1（MCP-1）的分泌，据报道MCP-1可增加细胞通透性，我们发现依达拉奉的一个新功能是部分通过下调MCP-1的分泌促进血管内皮细胞紧密连接的形成。这些数据为依达拉奉在脑血管疾病患者中的临床应用提供了基础和有用的信息。
Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a free radical scavenger used for acute ischemic stroke. However, it is not known whether edaravone works only as a free radical scavenger or possess other pharmacological actions. Therefore, we elucidated the effects of edaravone on human brain microvascular endothelial cells (HBMECs) by 2 dimensional fluorescence difference gel electrophoresis (2D-DIGE). We found 38 protein spots the intensity of which was significantly altered 1.3 fold on average (p< 0.05) by the edaravone treatment and successfully identified 17 proteins of those.Four of those 17 proteins were cytoskeleton proteins or cytoskeleton-regulating proteins. Therefore, we subsequently investigated the change of size and shape of the cells, the actin network, and the tight junction of HBMEC by immunocytochemistry. As a result, most edaravone-treated HBMECs became larger and rounder compared with those that were not treated. Furthermore, edaravone-treated HBMECs formed gathering zona occludens (ZO)-1, a tight junction protein, along the junction of the cells. In addition, we found that edaravone suppressed interleukin (IL)-1β-induced secretion of monocyte chemoattractant protein-1 (MCP-1), which was reported to increase cell permeability.We found a novel function of edaravone is the promotion of tight junction formations of vascular endothelial cells partly via the down-regulation of MCP-1 secretion. These data provide fundamental and useful information in the clinical use of edaravone in patients with cerebral vascular diseases.