Activation of the JAK-STAT signaling pathway in the rat basilar artery after subarachnoid hemorrhage
Activation of the JAK-STAT signaling pathway in the rat basilar artery after subarachnoid hemorrhage
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DOI：
10.1016/j.brainres.2005.12.003
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发表时间：
2006-02-09
期刊：
影响因子：
2.9
通讯作者：
Yoshida, J
中科院分区：
文献类型：
--
作者：
Osuka, K;Watanabe, Y;Yoshida, J
Janus激酶信号转导子和转录激活子（JAK-STAT）是细胞对细胞因子应答时从细胞表面转导信号的最重要的信号传导途径之一。蛛网膜下腔出血（SAH）在CSF中产生细胞因子。我们研究了SAH后大鼠基底动脉中细胞因子是否激活了该信号通路。在SAH大鼠单次出血模型中，直到SAH后7天才获得基底动脉和CSF。ELISA法测定脑脊液中白细胞介素6（IL-6）的含量。在基底动脉中进行JAK 1、磷酸化特异性JAK 1、STAT 3、Tyr（705）和Ser（727）处的磷酸化特异性STAT 3、环氧合酶-2（考克斯-2）和肌动蛋白抗体的Western印迹分析。应用免疫组化方法检测基底动脉中STAT 3、磷酸化STAT 3 Tye（705）和Ser（727）及考克斯-2的表达。SAH后IL-6的浓度立即增加，Western印迹分析显示JAK 1在2小时内磷酸化，伴随着STAT 3在Tyr（705）的磷酸化，在第1-2天延伸到Ser（727）。免疫组织化学显示STAT 3磷酸化发生在基底动脉的内皮细胞和平滑肌细胞。此外，脑池内注射IL-6本身显著增加了STAT 3在Tyr（705）和Ser（127）处的磷酸化。基底动脉内皮细胞考克斯-2的表达也上调。这些结果表明SAH在CSF中产生促炎细胞因子IL-6，其激活基底动脉中的JAK-STAT信号通路并诱导立即早期基因的转录。(c)2005 Elsevier B. V.保留所有权利。
The Janus kinase-signal transducer and activator of transcription (JAK-STAT) is one of the most important signaling pathways transducing signals from the cell surface in response to cytokines. Subarachnoid hemorrhage (SAH) produces cytokines in the CSF. We investigated whether this signaling pathway is activated in the rat basilar artery after SAH by cytokines. in a rat single-hemorrhage model of SAH, basilar arteries and CSF were obtained until 7 days after SAH. The concentration of interleukin-6 (IL-6) in CSF was measured by ELISA. Western blot analysis with JAK1, phosphospecific-JAK1, STAT3, phosphospecific STAT3 at Tyr(705) and Ser(727), cyclooxygenase-2 (COX-2), and actin antibodies was performed in basilar artery. The expressions of STAT3, phosphospecific STAT3 at Tye(705) and Ser(727), and COX-2 in basilar artery were examined by immunohistochemical studies. The concentration of IL-6 immediately increased after SAH and Western blot analysis revealed that JAK1 was phosphorylated within 2 h, accompanied by phosphorylation of STAT3 at Tyr(705), extending to Ser(727) at days 1-2. Immunohistochemistry revealed phosphorylation of STAT3 to occur in endothelial and smooth muscle cells of the basilar artery. In addition, intracisternal injection of IL-6 by itself significantly increased phosphorylation of STAT3 at Tyr(705) and Ser(127). Expression of COX-2 was also upregulated in endothelial cells of the basilar artery. These results indicate that SAH produces the proinflammatory cytokine IL-6 in the CSF, which activates the JAK-STAT signaling pathway in the basilar artery and induces transcription of immediate early genes. (c) 2005 Elsevier B.V. All rights reserved.