Liang Xiaotao, Xiong Yifan, Liu Xueqi, Liang Xiaoshan, Zhu Xiaoyu, Xie Wei
School of Traditional Chinese Medicine, Southern Medical University.
People's Hospital of Zhuhai High-tech Industrial Development Zone, Zhuhai 519000, Guangdong.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 May 20;45(5):986-994. doi: 10.12122/j.issn.1673-4254.2025.05.11.
To investigate the therapeutic mechanism of Granules (HXSFG) for alleviating central sensitization in a mouse model of chronic migraine (CM).
We analyzed the main chemical components of HXSFG through literature review and explored their pharmacological mechanisms by bioinformatics analyses. In a male C57BL/6J mouse model of CM established by intraperitoneal injections of nitroglycerin (10 mg/kg) every other day (5 injections), the effects of gavage with low, and high doses of HXSFG or intraperitoneal injections of topiramate for ameliorating central sensitization were evaluated using Von Frey test and a hot plate apparatus; the changes in expressions of inflammatory factors, the proteins in the TLR4/NF‑κB signaling pathway, and activation of c-Fos and CGRP were detected using RT-qPCR, Western blotting and immunofluorescence staining.
Network pharmacology analysis suggested that the main active components in HXSFG for alleviating CM included formononetin, paeoniflorin, quercetin, and tanshinone. Gene Ontology (GO) enrichment analysis identified 492 GO entries, comprising 366 biological processes, 46 cellular components, and 80 molecular functions. KEGG pathway enrichment analysis indicated that the Toll-like receptor and NF‑κB signaling pathways were crucial in mediating the therapeutic effects of HXSFG on CM. In the mouse models of CM, both topiramate and HXSFG treatments alleviated the symptoms of central sensitization, evidenced by improved mechanical and thermal pain thresholds in the mice. HXSFG significantly reduced the expression of c-Fos and CGRP, improved inflammatory markers, and downregulated the expressions of TLR4, p-NF‑κB, IL-1β, and TNF‑α proteins in the mouse models.
HXSFG effectively alleviates central sensitization in CM mice by modulating the inflammatory pathways and inhibiting the TLR4/ NF-κB signaling pathway, suggesting its potential as a therapeutic option for CM.
探讨化血生肌方颗粒（HXSFG）缓解慢性偏头痛（CM）小鼠模型中枢敏化的治疗机制。
通过文献回顾分析HXSFG的主要化学成分，并通过生物信息学分析探索其药理机制。在通过隔日腹腔注射硝酸甘油（10 mg/kg，共5次注射）建立的雄性C57BL/6J小鼠CM模型中，使用von Frey试验和热板仪评估灌胃低、高剂量HXSFG或腹腔注射托吡酯改善中枢敏化的效果；使用RT-qPCR、蛋白质免疫印迹法和免疫荧光染色检测炎症因子、TLR4/NF-κB信号通路中的蛋白质表达变化以及c-Fos和降钙素基因相关肽（CGRP）的激活情况。
网络药理学分析表明，HXSFG中缓解CM的主要活性成分包括芒柄花素、芍药苷、槲皮素和丹参酮。基因本体（GO）富集分析确定了492个GO条目，包括366个生物学过程、46个细胞成分和80个分子功能。京都基因与基因组百科全书（KEGG）通路富集分析表明，Toll样受体和NF-κB信号通路在介导HXSFG对CM的治疗作用中起关键作用。在CM小鼠模型中，托吡酯和HXSFG治疗均缓解了中枢敏化症状，表现为小鼠机械性和热痛阈值提高。HXSFG显著降低了CM小鼠模型中c-Fos和CGRP的表达，改善了炎症标志物，并下调了TLR4、磷酸化NF-κB、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）蛋白的表达。
HXSFG通过调节炎症通路和抑制TLR4/NF-κB信号通路有效缓解CM小鼠的中枢敏化，表明其作为CM治疗选择的潜力。
