Sensory Biology Unit, Translational Research Centre, Copenhagen University Hospital - Rigshospitalet, Glostrup, Denmark.
Section of Cell Biology and Physiology, Department of Biology, University of Copenhagen, Copenhagen, Denmark.
Headache. 2024 Jun;64(6):652-662. doi: 10.1111/head.14726. Epub 2024 May 3.
Migraine, a prevalent and debilitating disease, involves complex pathophysiology possibly including inflammation and heightened pain sensitivity. The current study utilized the complete Freund's adjuvant (CFA) model of inflammation, with onabotulinumtoxinA (BoNT/A) as a treatment of interest due to its use in clinical migraine management. Using an animal model, the study sought to investigate the role of BoNT/A in modulating CFA-induced inflammation, alterations in pain sensitivity, and the regulation of calcitonin gene-related peptide (CGRP) release. Further, we aimed to assess the changes in SNAP-25 through western blot analysis to gain insights into the mechanistic action of BoNT/A.
BoNT/A or control was administered subcutaneously at the periorbital region of rats 3 days before the induction of inflammation using CFA. Periorbital mechanical sensitivity was assessed post-inflammation, and alterations in CGRP release were evaluated. Changes in SNAP-25 levels were determined using western blot analysis.
Upon CFA-induced inflammation, there was a marked increase in periorbital mechanical sensitivity, with the inflammation side showing increased sensitivity compared to other periorbital areas. BoNT/A did decrease the withdrawal thresholds in the electronic von Frey test. Despite not being able to observe differences in pain thresholds or CGRP release, BoNT/A reduced baseline release under CFA inflamed conditions. Analysis of SNAP-25 levels in the trigeminal ganglion revealed both intact and cleaved forms that were notably elevated in BoNT/A-treated animals. These findings, derived from western blot analysis, suggest an effect on neurotransmitter release.
Our investigation highlights the role of BoNT/A in reducing baseline CGRP in the context of inflammation and its involvement in SNAP-25 cleavage. In contrast, BoNT/A did not appear to alter facial pain sensitivity induced by inflammation, suggesting that mechanisms other than baseline CGRP could be implicated in the elevated thresholds in the CFA model.
偏头痛是一种常见且使人虚弱的疾病,涉及复杂的病理生理学,可能包括炎症和疼痛敏感性增加。本研究利用完全弗氏佐剂(CFA)炎症模型,以肉毒毒素 A 型(BoNT/A)作为治疗药物,因为它在偏头痛的临床治疗中得到了应用。该研究使用动物模型,旨在探讨 BoNT/A 在调节 CFA 诱导的炎症、疼痛敏感性改变以及降钙素基因相关肽(CGRP)释放中的作用。此外,我们旨在通过 Western blot 分析评估 SNAP-25 的变化,以深入了解 BoNT/A 的作用机制。
在使用 CFA 诱导炎症前 3 天,将 BoNT/A 或对照药物皮下注射到大鼠的眶周区域。在炎症后评估眶周机械敏感性,并评估 CGRP 释放的变化。通过 Western blot 分析确定 SNAP-25 水平的变化。
在 CFA 诱导的炎症中,眶周机械敏感性明显增加,炎症侧的敏感性高于其他眶周区域。BoNT/A 确实降低了电子von Frey 测试中的退缩阈值。尽管在疼痛阈值或 CGRP 释放方面未能观察到差异,但 BoNT/A 减少了 CFA 炎症条件下的基础释放。三叉神经节中 SNAP-25 水平的分析显示,在 BoNT/A 处理的动物中,既有完整形式,也有切割形式,均显著升高。这些来自 Western blot 分析的发现表明,BoNT/A 对神经递质释放有影响。
我们的研究强调了 BoNT/A 在减少炎症背景下基础 CGRP 释放方面的作用,以及其在 SNAP-25 切割中的参与。相比之下,BoNT/A 似乎并没有改变炎症引起的面部疼痛敏感性,这表明在 CFA 模型中,升高的阈值可能涉及除基础 CGRP 之外的其他机制。