Guo Guangyu, Zhang Lei, Liu Xuyang, Deng Yiping, Wu Peiyu, Zhao Ruofan, Wang Wei
Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Clinical Systems Biology Laboratories, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
J Headache Pain. 2025 May 26;26(1):130. doi: 10.1186/s10194-025-02058-4.
Migraine, characterized by recurrent episodes of severe headache, remains mechanistically enigmatic. While traditional theories emphasize trigeminovascular activation, the role of meningeal stromal-immune crosstalk in disease chronicity is poorly understood.
A migraine-related chronic hypersensitivity model was utilized via intermittent intraperitoneal nitroglycerin (NTG, 10 mg/kg, every other day for 9 days) and peripheral mechanical hypersensitivity was assessed using von Frey filaments. Single-cell RNA sequencing (scRNA-seq) was performed on dura tissues to construct a cellular atlas of NTG-induced remodeling. These data were then integrated with migraine genome-wide association study (GWAS) risk genes, cell-cell interaction networks, and transcriptional regulation analysis to dissect NTG-driven meningeal remodeling.
The NTG-induced migraine-related chronic hypersensitivity model demonstrated sustained mechanical allodynia, as evidenced by significantly decreased paw withdrawal thresholds (p < 0.0001). Single-cell profiling of the dura mater revealed a 2.4-fold expansion of a pro-inflammatory fibroblast subpopulation (Fibro_c5: 1.9% in Vehicle vs. 4.6% in NTG group), which exhibited marked activation of TNF-α/NF-κB signaling pathways (normalized enrichment score [NES] = 1.83). Concomitantly, we observed an 82% increase in meningeal monocytes (5.7-10.4%) that showed preferential interaction with Fibro_c5 fibroblasts through Angptl1-mediated stromal-immune crosstalk (log2 fold change = 1.41). Regulatory network analysis identified Mafk as the upstream transcriptional regulator orchestrating Angptl1 expression in this pathological communication axis.
Our study reveals that NTG reprograms meningeal fibroblasts to expand a pro-inflammatory fibroblast subtype, which drives migraine-related chronic hypersensitivity through TNF-α/NF-κB signaling and Angptl1-mediated monocyte crosstalk. The identified Mafk-Angptl1 axis presents a potential therapeutic target, though human validation remains essential.
偏头痛以反复发作的严重头痛为特征，其发病机制仍不明确。虽然传统理论强调三叉神经血管激活，但脑膜基质-免疫相互作用在疾病慢性化中的作用却知之甚少。
通过间歇性腹腔注射硝酸甘油（NTG，10mg/kg，隔日一次，共9天）建立偏头痛相关慢性超敏反应模型，并使用von Frey细丝评估外周机械性超敏反应。对硬脑膜组织进行单细胞RNA测序（scRNA-seq），以构建NTG诱导重塑的细胞图谱。然后将这些数据与偏头痛全基因组关联研究（GWAS）风险基因、细胞-细胞相互作用网络和转录调控分析相结合，以剖析NTG驱动的脑膜重塑。
NTG诱导的偏头痛相关慢性超敏反应模型表现出持续的机械性痛觉过敏，爪部撤离阈值显著降低证明了这一点（p<0.0001）。硬脑膜的单细胞分析显示，促炎成纤维细胞亚群（Fibro_c5：对照组为1.9%，NTG组为4.6%）扩大了2.4倍，该亚群表现出TNF-α/NF-κB信号通路的显著激活（标准化富集分数[NES]=1.83）。同时，我们观察到脑膜单核细胞增加了82%（从5.7%增至10.4%），这些单核细胞通过血管生成素样蛋白1（Angptl1）介导的基质-免疫相互作用与Fibro_c5成纤维细胞优先相互作用（log2倍数变化=1.41）。调控网络分析确定Mafk是在这个病理通讯轴中协调Angptl1表达的上游转录调节因子。
我们的研究表明，NTG对脑膜成纤维细胞进行重编程，以扩大一种促炎成纤维细胞亚型，该亚型通过TNF-α/NF-κB信号通路和Angptl1介导的单核细胞相互作用驱动偏头痛相关的慢性超敏反应。虽然仍需人体验证，但已确定的Mafk-Angptl1轴是一个潜在的治疗靶点。
