Preventive treatment of nitroglycerin-induced chronic migraine by peripheral single injection of Botulinum Neurotoxin A in mice
10.3760/cma.j.cn115354-20210621-00391
- VernacularTitle:A型肉毒毒素预防小鼠慢性偏头痛发作的机制研究
- Author:
Ting ZHU
1
;
Jingqi NIU
;
Cunjin SU
;
Weijia CHEN
;
Yanlin ZHANG
;
Chunfeng LIU
;
Tong LIU
;
Weifeng LUO
Author Information
1. 苏州大学附属第二医院神经内科，苏州 215004
- Keywords:
Chronic migraine;
Botulinum neurotoxin A;
Calcitonin gene-related peptide;
NOD-like receptor protein 3
- From:
Chinese Journal of Neuromedicine
2022;21(5):433-442
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the effect of botulinum neurotoxin A (BoNT/A) on prevention of chronic migraine (CM) in mice and explore the potential mechanism.Methods:Twenty-four male C57BL/6 mice were randomly divided into control group, nitroglycerin (NTG) group, and BoNT/A+NTG group ( n=8). Mice in the latter two groups were intraperitoneally injected with 10 mg/kg NTG on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments to establish CM models. Mice in the BoNT/A+NTG group were injected with 0.18 U/100 μL BoNT/A one h before the first injection of NTG. Mice in the control group were injected with the same dose of normal saline. Basal mechanical withdrawal threshold (MWT) and evoked MWT 2 h after NTG in the facial and hindpaw regions on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments were evaluated by von Frey filament test. The motor function of mice 2 h after NTG injection was tested by rotarod test on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments. On 9 th d of experiments, the mice were sacrified; the calcitonin gene-related peptide (CGRP), synaptosomal-associated protein 25 (SNAP25), glial fibrillary acidic protein (GFAP) and TRP channel protein expressions in the trigeminal ganglia (TG) and trigeminal nucleus caudalis (TNC), and NOD-like receptor protein 3 (NLRP3) inflammatory factor pathway-related protein expressions in TNC were detected by Western blotting; real-time quantitative PCR (RT-qPCR) was used to detect the NLRP3 inflammatory factor pathway-related mRNA expressions in TNC. The CGRP expression in TNC was detected by immunofluorescent staining. Results:(1) As compared with the control group, the NTG group had significantly decreased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05). (2) There was no significant difference in running time on rotarod among the three groups ( P>0.05). (3)Western blotting results showed that as compared with those in the control group, the CGRP and SNAP25 protein expressions were significantly increased in TG of the NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). As compared with those in the control group, the CGRP and NLRP3 protein expressions were significantly increased in TNC of NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). (4)RT-qPCR results showed that as compared with that in the control group, the IL-1β mRNA expression in TNC of the NTG group was significantly increased ( P<0.05), and that in the BoNT/A prevention group was statistically decreased as compared with that in the NTG group ( P<0.05). (5) Immunofluorescent staining results showed that as compared with that in the control group, the CGRP expression in TNC of the NTG group was significantly increased, and that in the BoNT/A+NTG group was significantly decreased as compared with that in the NTG group ( P<0.05). Conclusion:BoNT/A can reduce the SNAP25 expression in TG, reduce the CGRP release in TG and TNC, and prevent CM onset; BoNT/A can regulate NLRP3 level in TNC.
