Botulinum Neurotoxin A Prevents the Development of Nitroglycerin-Induced Chronic Migraine via Inhibition of CGRP and NLRP3 Inflammasomes in Mice

Abstract

Background This study was designed to examine the therapeutic effects of Botulinum neurotoxin A (BoNT/A) on chronic migraine and to explore the potential mechanisms by using a mouse model of chronic migraine, which was established by repeated intraperitoneal (i.p.) injection with nitroglycerin (NTG).Methods NTG-induced basal and evoked mechanical hypersensitivity were evaluated using the von Frey filament test. Before the first injection of NTG, a single facial injection of BoNT/A was administered in the supraorbital region to explore its preventive effects on the development of chronic migraine in mice. The expression of calcitonin gene-related peptide (CGRP) and synaptosomal-associated protein25 (SNAP25) in the trigeminal ganglia (TG) and the trigeminal nucleus caudalis (TNC) were detected by Western blotting and immunostaining. Results Repeated administration of NTG resulted in basal and evoked mechanical hypersensitivity in mice. Single facial BoNT/A injection prevented the development of NTG-induced mechanical hypersensitivity in mice. Western blotting results revealed that peripheral BoNT/A injection decreased the NTG-induced upregulation of expression of CGRP and SNAP25 in the TGs and NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasomes in the TNC. Immunostaining results revealed peripheral BoNT/A injection also decreased the NTG-induced upregulation of CGRP expression in the TNC.Conclusions Thus, these results indicate that single facial injection of BoNT/A may be a preventive treatment of chronic migraine, possible due to the inhibitory effect of BoNT/A on the expression of CGRP and SNAP25 in the TGs and NLRP3 inflammasomes in the TNC.

A role of NLRP3 and MMP9 in migraine progression: a systematic review of translational study

“…Across the research included in this review, a variety of outcome measures were observed, such as behavioral tests ( n = 9; 40.09%) ( 30 , 70 , 74–76 ), Sensory sensitivity testing ( 28 ), von Frey Testing ( 27 , 77 ), Light/dark test ( 27 ), Von frey test ( 27 ). To check the specific biomarker estimation ELISA ( n = 8; 36.36%) ( 30 , 31 , 72 , 74 , 75 , 77–79 ), Western blot analysis ( n = 9; 40.09%) ( 28 , 70 , 73–75 , 79–83 ), immunohistochemistry ( n = 4; 18.18%) ( 72 , 73 , 75 , 77 , 78 , 81 , 82 ), Histopathology ( 27 ), Immunofuorescence staining ( n = 9; 40.09%) ( 27 , 28 , 30 , 31 , 70 , 76 , 78 ), and for gene expression used qRT PCR ( n = 7; 31.81%) ( 27 , 28 , 31 , 70 , 74 , 77 , 78 , 80 , 81 , 83 ). The majority of researchers, around 60–70%, neglect to validate their experiments using behavioral tests, Western blotting, Immunofluorescence, and Immunohistochemistry, despite these tests being crucial for experimental validation.…”

“…Upon activation of the Trigeminal nerve system pathway, microglia release an array of chemokines, cytokines, as well as free radicals including nitric oxide and reactive oxygen species. Nevertheless, further investigation into brain microglia is necessary to advance therapy ( 76 ). Excessively active microglia can potentially lead to neuroinflammation and tissue damage.…”

“…, Immunofuorescence staining (n = 9; 40.09%)(27, 28,30,31,70,76,78), and for gene expression used qRT PCR (n = 7; 31.81%)(27, 28,31,70, 74, 77,78, 80,81,83). The majority of researchers, around 60-70%, neglect to validate their experiments using behavioral tests, Western blotting, Immunofluorescence, and Immunohistochemistry, despite these tests being crucial for experimental validation.…”

Botulinum toxin type A ameliorates rat dorsal root ganglia neuron pyroptosis in postherpetic neuralgia by upregulating cathelicidin antimicrobial peptide to inhibit neutrophil elastase

“…However, the mechanism of BoNT/A on PHN remains elusive. In a chronic migraine mice model, BoNT/A injection downregulates NLRP3 and IL‐1β expression, indicating its inhibitory effect on NLRP3 inflammasome activation (T. Zhu et al., 2020). In this case, BoNT/A may ameliorate PHN via regulating NLRP3 inflammasome‐induced pyroptosis.…”