Role of SCH79797 in maintaining vascular integrity in rat model of subarachnoid hemorrhage.
Role of SCH79797 in maintaining vascular integrity in rat model of subarachnoid hemorrhage.
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DOI：
10.1161/strokeaha.113.678474
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发表时间：
2013-05
期刊：
影响因子：
8.3
通讯作者：
Zhang JH
中科院分区：
文献类型：
--
作者：
Yan J;Manaenko A;Chen S;Klebe D;Ma Q;Caner B;Fujii M;Zhou C;Zhang JH
蛛网膜下腔出血 (SAH) 后血浆凝血酶浓度升高。然而，凝血酶受体（蛋白酶激活受体-1，PAR-1）在内皮屏障破坏中的作用尚未研究。本研究的目的是调查 PAR-1 在调节血管通透性中的作用，并评估 PAR-1 拮抗剂 SCH79797 通过维持血管完整性的潜在治疗作用。通过血管内穿孔将 SCH79797 注射到经历 SAH 的雄性 Sprauge-Dawley 大鼠腹腔内。 SAH后24小时评估脑含水量、伊文思蓝含量和神经行为测试。为了探讨SAH后PAR-1激活的作用以及SCH79797作用的具体机制，我们对海马组织进行了Western blot、免疫沉淀和免疫荧光检测。使用 p21 激活激酶 1 (PAK1) 抑制剂 IPA-3 来探索 SCH79797 的潜在保护机制。 SAH 后 24 小时，接受 SCH79797 治疗的动物表现出脑含水量、伊文思蓝含量和神经行为缺陷的减少。 SCH79797 还减弱 PAR-1 表达并维持 VE-钙粘蛋白（粘附连接的重要组成部分）的水平。在 PAR-1 下游，PAK1 的 c-Src 依赖性激活导致 VE-钙粘蛋白的丝氨酸/苏氨酸磷酸化增加；免疫沉淀结果显示磷酸化 VE-钙粘蛋白与内吞作用协调器 β-arrestin2 的结合增强。 SCH79797 治疗后这些病理状态得到抑制。 SAH 后 PAR-1 激活至少部分通过 PAR-1-c-Src-PAK1-VE-钙粘蛋白磷酸化途径增加微血管通透性。通过抑制 PAR-1 活性，SCH79797 在 SAH 后维持微血管完整性方面发挥保护作用。
Plasma thrombin concentration is increased following subarachnoid hemorrhage (SAH). However, the role of thrombin receptor (protease activated recptor-1, PAR-1) in endothelial barrier disruption has not been studied. The aims of this study were to investigate the role of PAR-1 in orchestrating vascular permeability and assess the potential therapeutics of a PAR-1 antagonist, SCH79797, through maintaining vascular integrity. SCH79797 was injected intraperitoneally into male Sprauge-Dawley rats undergoing SAH by endovascular perforation. Assessment was conducted at 24 hours after SAH for brain water content, Evans blue content, and neurobehavioral testing. To explore the role of PAR-1 activation and the specific mechanism of SCH79797’s effect after SAH, Western blot, immunoprecipitation, and immunofluorescence of hippocampus tissue were performed. A p21-activated kinase1 (PAK1) inhibitor, IPA-3, was used to explore the underlying protective mechanism of SCH79797. At 24 hours after SAH, animals treated with SCH79797 demonstrated a reduction in brain water content, Evans blue content, and neurobehavioral deficits. SCH79797 also attenuated PAR-1 expression and maintained the level of VE-cadherin, an important component of adherens junctions. Downstream to PAR-1, c-Src dependent activation of PAK1 led to an increased serine/threonine phosphorylation of VE-cadherin; immunoprecipitation results revealed an enhanced binding of phosphorylated VE-cadherin with endocytosis orchestrator β-arrestin2. These pathological states were suppressed following SCH79797 treatment. PAR-1 activation following SAH increases microvascular permeability, at least, partly through a PAR-1-c-Src-PAK1-VE-cadherin phosphorylation pathway. Through suppressing PAR-1 activity, SCH79797 plays a protective role in maintaining microvascular integrity after SAH.