TRAUMATIC INJURY ELICITS JNK-MEDIATED HUMAN ASTROCYTE RETRACTION IN VITRO
TRAUMATIC INJURY ELICITS JNK-MEDIATED HUMAN ASTROCYTE RETRACTION IN VITRO
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DOI：
10.1016/j.neuroscience.2014.05.009
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发表时间：
2014-08-22
期刊：
影响因子：
3.3
通讯作者：
Fraser, D. D.
中科院分区：
文献类型：
--
作者：
Augustine, C.;Cepinskas, G.;Fraser, D. D.
脑损伤导致血脑屏障（BBB）功能障碍。血脑屏障由血管周围的星形胶质细胞组成，星形胶质细胞的终足包裹脑微血管内皮细胞。我们研究了体外创伤诱导的人星形胶质细胞（HA）和人脑微血管内皮细胞（hCMEC/D3）的形态学变化，包括丝裂原活化蛋白激酶（MAPK）信号转导通路的潜在作用。HA或hCMEC/D3在柔性培养膜上生长，并经受标准化为20%、30%或55%膜变形的单一创伤性损伤。测定细胞的形态学变化（即收缩）和MAPK磷酸化和/或表达（c-Jun NH2-末端激酶（JNK）1/2、细胞外信号调节激酶（ERK）1/2和p38）。单次创伤性损伤可迅速引起HA回缩（55%膜变形; p <0.01）。2 h内观察到HA形态学恢复（p <0.05）。创伤性损伤增加HA中磷酸化JNK1/2（p <0.05），表明MAPK活化。用结构不同的JNK抑制剂（25 μ M），SP 600125或SU 3327预处理HA，减少JNK磷酸化（p <0.05）和创伤诱导的HA收缩（P <0.05）。与HA相反，创伤性损伤未能诱导hCMEC/D3的形态学变化或MAPK活化。总之，创伤性损伤诱导JNK介导的HA在体外收缩，同时保留脑微血管内皮细胞的形态学变化。脑外伤后可发生星形胶质细胞从微血管内皮细胞回缩，导致细胞解偶联和血脑屏障功能障碍。JNK可能是创伤性脑损伤的潜在治疗靶点。(C)2014年IBRO。由爱思唯尔有限公司出版。保留所有权利。
Brain injury causes dysfunction of the blood-brain barrier (BBB). The BBB is comprised of perivascular astrocytes whose end-feet ensheath brain microvascular endothelial cells. We investigated trauma-induced morphological changes of human astrocytes (HA) and human cerebral microvascular endothelial cells (hCMEC/D3) in vitro, including the potential role of mitogen-activated protein kinase (MAPK) signal-transduction pathways. HA or hCMEC/D3 were grown on flexible culture membranes and subjected to single traumatic injury normalized to 20%, 30% or 55% membrane deformation. Cells were assayed for morphological changes (i.e. retraction) and MAPK phosphorylation and/or expression (c-Jun NH2-terminal kinase (JNK) 1/2, extracellular signal-regulated kinase (ERK) 1/2, and p38). HA retraction was rapidly elicited with a single traumatic injury (55% membrane deformation; p < 0.01). Morphological recovery of HA was observed within 2 h (p < 0.05). Traumatic injuries increased phospho-JNK1/2 (p < 0.05) in HA, indicating MAPK activation. Pre-treatment of HA with structurally distinct JNK inhibitors (25 mu M), either SP600125 or SU3327, reduced JNK phosphorylation (p < 0.05) and trauma-induced HA retraction (P < 0.05). In contrast to HA, traumatic injury failed to induce either morphological changes or MAPK activation in hCMEC/D3. In summary, traumatic injury induces JNK-mediated HA retraction in vitro, while sparing morphological changes in cerebral microvascular endothelial cells. Astrocyte retraction from microvascular endothelial cells in vivo may occur after brain trauma, resulting in cellular uncoupling and BBB dysfunction. JNK may represent a potential therapeutic target for traumatic brain injuries. (C) 2014 IBRO. Published by Elsevier Ltd. All rights reserved.