Department of Neurology, the First Medical Center, Chinese PLA General Hospital, Fuxing Road 28, Haidian District, Beijing, 100853, China.
Medical School of Chinese PLA, Beijing, 100853, China.
J Headache Pain. 2024 Apr 2;25(1):50. doi: 10.1186/s10194-024-01750-1.
Migraine stands as a prevalent primary headache disorder, with prior research highlighting the significant involvement of oxidative stress and inflammatory pathways in its pathogenesis and chronicity. Existing evidence indicates the capacity of Dl-3-n-butylphthalide (NBP) to mitigate oxidative stress and inflammation, thereby conferring neuroprotective benefits in many central nervous system diseases. However, the specific therapeutic implications of NBP in the context of migraine remain to be elucidated.
We established a C57BL/6 mouse model of chronic migraine (CM) using recurrent intraperitoneal injections of nitroglycerin (NTG, 10 mg/kg), and prophylactic treatment was simulated by administering NBP (30 mg/kg, 60 mg/kg, 120 mg/kg) by gavage prior to each NTG injection. Mechanical threshold was assessed using von Frey fibers, and photophobia and anxious behaviours were assessed using a light/dark box and elevated plus maze. Expression of c-Fos, calcitonin gene-related peptide (CGRP), Nucleus factor erythroid 2-related factor 2 (Nrf2) and related pathway proteins in the spinal trigeminal nucleus caudalis (SP5C) were detected by Western blotting (WB) or immunofluorescence (IF). The expression of IL-1β, IL-6, TNF-α, Superoxide dismutase (SOD) and malondialdehyde (MDA) in SP5C and CGRP in plasma were detected by ELISA. A reactive oxygen species (ROS) probe was used to detect the expression of ROS in the SP5C.
At the end of the modelling period, chronic migraine mice showed significantly reduced mechanical nociceptive thresholds, as well as photophobic and anxious behaviours. Pretreatment with NBP attenuated nociceptive sensitization, photophobia, and anxiety in the model mice, reduced expression levels of c-Fos and CGRP in the SP5C and activated Nrf2 and its downstream proteins HO-1 and NQO-1. By measuring the associated cytokines, we also found that NBP reduced levels of oxidative stress and inflammation. Most importantly, the therapeutic effect of NBP was significantly reduced after the administration of ML385 to inhibit Nrf2.
Our data suggest that NBP may alleviate migraine by activating the Nrf2 pathway to reduce oxidative stress and inflammation in migraine mouse models, confirming that it may be a potential drug for the treatment of migraine.
偏头痛是一种常见的原发性头痛疾病，先前的研究强调了氧化应激和炎症途径在其发病机制和慢性化过程中的重要作用。现有证据表明，Dl-3-正丁基苯酞（NBP）具有减轻氧化应激和炎症的能力，从而在许多中枢神经系统疾病中具有神经保护作用。然而，NBP 在偏头痛中的具体治疗意义仍有待阐明。
我们使用反复腹腔内注射硝化甘油（NTG，10mg/kg）建立了 C57BL/6 慢性偏头痛（CM）小鼠模型，并通过在每次 NTG 注射前给予 NBP（30mg/kg、60mg/kg、120mg/kg）灌胃模拟预防性治疗。使用 von Frey 纤维评估机械阈值，使用光/暗箱和高架十字迷宫评估畏光和焦虑行为。通过 Western blot（WB）或免疫荧光（IF）检测脊髓三叉神经核尾侧（SP5C）中 c-Fos、降钙素基因相关肽（CGRP）、核因子红细胞 2 相关因子 2（Nrf2）和相关通路蛋白的表达。通过 ELISA 检测 SP5C 中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、超氧化物歧化酶（SOD）和丙二醛（MDA）的表达以及血浆中 CGRP 的表达。使用活性氧（ROS）探针检测 SP5C 中 ROS 的表达。
在建模期末，慢性偏头痛小鼠的机械痛觉敏化阈值显著降低，且出现畏光和焦虑行为。NBP 预处理可减轻模型小鼠的痛觉敏化、畏光和焦虑，降低 SP5C 中 c-Fos 和 CGRP 的表达水平，并激活 Nrf2 及其下游蛋白 HO-1 和 NQO-1。通过测量相关细胞因子，我们还发现 NBP 降低了氧化应激和炎症水平。最重要的是，用 ML385 抑制 Nrf2 后，NBP 的治疗效果明显降低。
我们的数据表明，NBP 可能通过激活 Nrf2 通路减轻偏头痛小鼠模型中的氧化应激和炎症，从而缓解偏头痛，证实其可能成为治疗偏头痛的潜在药物。
