Reuter U, Bolay H, Jansen-Olesen I, Chiarugi A, Sanchez del Rio M, Letourneau R, Theoharides T C, Waeber C, Moskowitz M A
Stroke and Neurovascular Regulation Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA.
Brain. 2001 Dec;124(Pt 12):2490-502. doi: 10.1093/brain/124.12.2490.
Nitric oxide (NO) has been implicated in migraine pathogenesis based on the delayed development of typical migraine headache 4-6 h after infusing the NO donor nitroglycerin [glyceryl trinitrate (GTN)] to migraineurs. Furthermore, inhibiting the synthesis of NO by treatment with a NO synthase (NOS) inhibitor attenuates spontaneous migraine headaches in 67% of subjects. Because NO has been linked to inflammation and cytokine expression, we investigated the delayed consequences of brief GTN infusion (30 min) on the development of meningeal inflammation in a rat model using doses relevant to the human model. We found dose-dependent Type II NOS [inducible NOS (iNOS)] mRNA upregulation in dura mater beginning at 2 h and an increase in the corresponding protein expression at 4, 6 and 10 h after infusion. Type II NOS immunoreactivity was expressed chiefly within resident meningeal macrophages. Consistent with development of a delayed inflammatory response, we detected induction of interleukin 1beta in dura mater at 2 and 6 h and increased interleukin 6 in dural macrophages and in rat cerebrospinal fluid at 6 h after GTN infusion. Myeloperoxidase-positive cells were rarely found. Leakage of plasma proteins from dural blood vessels was first detected 4 h after GTN infusion, and this was suppressed by administering a specific Type II NOS inhibitor [L-N(6)-(1-iminoethyl)-lysine (L-NIL)]. In addition to cytokine induction, macrophage iNOS upregulation and oedema formation after GTN infusion, dural mast cells exhibited granular changes consistent with secretion at 4 and 6 h. Because iNOS was expressed in dural macrophages following topical GTN, and in the spleen after intravenous injection, the data suggest that the inflammatory response is mediated by direct actions on the dura and does not develop secondary to events within the brain. Our findings point to the importance of new gene expression and cytokine expression as fundamental to the delayed response following GTN infusion, and support the hypothesis that a similar response develops in human meninges after GTN challenge.
基于向偏头痛患者输注一氧化氮（NO）供体硝酸甘油[三硝酸甘油酯（GTN）]后4-6小时典型偏头痛头痛延迟发作，NO已被认为与偏头痛发病机制有关。此外，用一氧化氮合酶（NOS）抑制剂治疗抑制NO合成可使67%的受试者自发性偏头痛头痛减轻。由于NO与炎症和细胞因子表达有关，我们在大鼠模型中使用与人模型相关的剂量，研究了短暂输注GTN（30分钟）对脑膜炎症发展的延迟影响。我们发现，输注后2小时硬脑膜中II型NOS[诱导型NOS（iNOS）]mRNA呈剂量依赖性上调，4、6和10小时相应蛋白表达增加。II型NOS免疫反应主要在驻留脑膜巨噬细胞内表达。与延迟炎症反应的发展一致，我们在GTN输注后2和6小时在硬脑膜中检测到白细胞介素1β的诱导，6小时在硬脑膜巨噬细胞和大鼠脑脊液中检测到白细胞介素6增加。很少发现髓过氧化物酶阳性细胞。GTN输注后4小时首次检测到硬脑膜血管血浆蛋白渗漏，施用特异性II型NOS抑制剂[L-N（6）-（1-亚氨基乙基）-赖氨酸（L-NIL）]可抑制这种渗漏。除了GTN输注后细胞因子诱导、巨噬细胞iNOS上调和水肿形成外，硬脑膜肥大细胞在4和6小时表现出与分泌一致的颗粒变化。由于局部应用GTN后iNOS在硬脑膜巨噬细胞中表达，静脉注射后在脾脏中表达，数据表明炎症反应是由对硬脑膜的直接作用介导的，并非继发于脑内事件。我们的研究结果指出新基因表达和细胞因子表达对于GTN输注后延迟反应至关重要，并支持这样的假说，即GTN激发后人类脑膜会发生类似反应。
