瞬时受体电位通道家族M8亚型在大鼠偏头痛发病机制中的作用

秦冬梅; 邹撰; 周超然; 母发光

doi:10.7499/j.issn.1008-8830.2015.05.020

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中国当代儿科杂志 ›› 2015, Vol. 17 ›› Issue (5) : 515-519. DOI: 10.7499/j.issn.1008-8830.2015.05.020

论著·实验研究

瞬时受体电位通道家族M8亚型在大鼠偏头痛发病机制中的作用

秦冬梅, 邹撰, 周超然, 母发光

作者信息 +

Role of transient receptor potential melastatin 8 channels in migraine mechanism in rats

QIN Dong-Mei, ZOU Zhuan, ZHOU Chao-Ran, MU Fa-Guang

Author information +

文章历史 +

摘要

目的　通过检测瞬时受体电位通道家族M8(TRPM8)亚型在偏头痛模型大鼠三叉神经组织中的表达变化探讨TRPM8在大鼠偏头痛发病机制中的作用。方法　将20只雄性Sprague-Dawley(SD)大鼠随机分为空白对照组和模型组, 每组10只。采用颈背部每周1次皮下注射硝酸甘油(10 mg/kg)建立无先兆性偏头痛大鼠模型, 共5周; 对照组则用生理盐水代替硝酸甘油行皮下注射。最后一次皮下注射4 h后, 对各组大鼠行行为学评分, 而后采集两组大鼠颅底三叉神经节, 采用免疫组化法检测两组三叉神经节中谷氨酸受体(NMDAR)的表达变化; 采用免疫荧光检测法观察两组三叉神经节中蛋白激酶A(PKA)的表达; 采用western blot法检测TRPM8蛋白在两组中的表达情况。结果　模型组大鼠在建模过程中每周的行为学评分均高于对照组(P<0.05)。模型组中NMDAR、PKA及TRPM8表达水平均高于对照组(P<0.01)。行为学评分与TRPM8表达水平呈正相关(r=0.822, P<0.01); NMDAR与TRPM8表达水平亦呈正相关(r=0.794, P<0.01)。结论 TRPM8可能参与了偏头痛的发病机制, 并且可能是通过NMDAR途径实现。

Abstract

Objective To investigate the role of transient receptor potential melastatin 8 (TRPM8) channels in migraine mechanism in rats by measuring the changes in expression of TRPM8 in the trigeminal nerve of rats with migraine. Methods Twenty male Sprague-Dawley rats were randomly and equally divided into a blank control group and a model group. Nitroglycerin (10 mg/kg) was injected subcutaneously in the back of the neck once a week for 5 weeks, to prepared a rat model of migraine without aura. Normal saline was injected subcutaneously instead of nitroglycerin in the control group. At 4 hours after the final injection, behavior scoring of all rats was performed, and then the trigeminal nerve ganglions of rats in both groups were collected for measurement of expression of N-methyl-D-aspartate receptor (NMDAR), protein kinase A (PKA), and TRPM8 using immunohistochemical staining, immunofluorescence, and Western blot, respectively. Results The behavior score in each week during the rat model preparing was significantly higher in the model group than in the control group (P<0.05). The expression of NMDAR, PKA, and TRPM8 in the model group was significantly higher than in the control group (P<0.01). Both the behavior score and the expression of NMDAR were positively correlated with the expression of TRPM8 (r=0.822 and 0.794 respectively; P<0.01). Conclusions TRPM8 may be involved in migraine mechanism probably by activation of the NMDAR pathway.

导出引用

秦冬梅, 邹撰, 周超然, 母发光. 瞬时受体电位通道家族M8亚型在大鼠偏头痛发病机制中的作用[J]. 中国当代儿科杂志. 2015, 17(5): 515-519 https://doi.org/10.7499/j.issn.1008-8830.2015.05.020

QIN Dong-Mei, ZOU Zhuan, ZHOU Chao-Ran, MU Fa-Guang. Role of transient receptor potential melastatin 8 channels in migraine mechanism in rats[J]. Chinese Journal of Contemporary Pediatrics. 2015, 17(5): 515-519 https://doi.org/10.7499/j.issn.1008-8830.2015.05.020

参考文献

[1] Soon YY, Siow HC, Tan CY. Assessment of migraineur referred to a specialist headache clinic in Singapore: diagnosis, treatment strategies, outcomes, knowledge of migraine treatments and satisfaction[J]. Cephalalgia, 2005, 25(1): 122-132.
[2] 方洁, 王子才. 托吡酯治疗儿童偏头痛的临床研究[J]. 中国当代儿科杂志, 2006, 8(6): 509-510.
[3] Xiao Y, Richter JA, Hurley JH. Release of glutamate and CGRP from trigeminal ganglion neurons: Role of calcium channels and 5-HT1 receptor signaling[J]. Moll Pain, 2008, 4: 12.
[4] Chasman DI, Schurks M, Anttila V, et al. Genome-wide association study reveals three susceptibility loci for common migraine in the general population[J]. Nat Genet, 2011, 12(7): 695-843.
[5] Markus Schurks. Genetics of migraine in the age of genome-wide association studies[J]. J Headache Pain, 2012, 13(1): 1-9.
[6] Greg A, Zameel M. New directions in migraine[J]. Weir and Cader BMC Medicine, 2011, 9: 116.
[7] Devesa I, Ferrer-Montiel A. Neurotrophins, endocannabinoids and thermo-transient receptor potential: a threesome in pain signaling[J]. Eur J Neurosci, 2014, 39(3): 353-362.
[8] Tassorelli C, Greco R, Morocutti A, et al. Nitricoxide-induced neuronal activation in the central nervous system as an animal model of migraine: mechanisms and mediators[J]. Funct Neurol, 2001, 16(4 Suppl): 69-76.
[9] Sarria I, Gu J. Menthol response and adaptation in nociceptive-like and nonnociceptive-like neurons: role of protein kinases[J]. Moll Pain, 2010, 6: 47.
[10] 朱晓凤, 韩月臣, 樊兆民, 等. 热凝脑膜中动脉对硝酸甘油致偏头痛大鼠三叉神经尾核c-foes基因表达的影响[J]. 山东大学耳鼻喉眼学报, 2010, 24(1): 61-64.
[11] 杨丽萍, 张育才. 儿童偏头痛的临床研究进展[J]. 临床儿科杂志, 2005, 23(4): 246-247.
[12] 吴希如, 林庆. 小儿神经系统疾病基础与临床[M]. 北京: 人民出版社, 2000: 633-637.
[13] 陈锦泳, 尹昭. 儿童和青少年偏头痛[J]. 新医学杂志, 2009, 40(12): 771-774.
[14] 郭琳, 洪治平. 硝酸甘油型实验性偏头痛模型原理与研究现状[J]. 中国疼痛医学杂志, 2004, 10(6): 357-364.
[15] 吴敏, 陈美娟. 偏头痛发病机制的研究进展[J]. 四川生理科学杂志, 2007, 29(4): 173-176.
[16] 张敏. 偏头痛发病机制的研究进展[J]. 中国临床医生, 2010, 38(9): 14-15.
[17] 姚宝珍, 丁琼, 凌伟, 等. 儿童偏头痛的临床特点及脑电图表现[J]. 中国当代儿科杂志, 2002, 4(5): 395-396.
[18] 郭张华, 陈建平. NMDA 受体在神经病理性疼痛中作用的研究进展[J]. 中国疼痛医学杂志, 2013, 19(3): 173-176.
[19] Proudfoot CJ, Garry EM, Cottrell DF, et al. Analgesia mediated by the TRPM8 cold receptor in chronic neuropathic pain[J]. Curr Biol, 2006, 16(16): 1591-1605.
[20] Kayo T, Kempski O, Hermann A, et al. Cluster analysis of mRNA expression levels identifies multiple sequential patterns following focal cerebral ischemia[J]. Turk Neuron Surge, 2012, 22(4): 441-447.
[21] 朱正萍, 杨晓苏. PKC-γ/NMDAR1在硝酸甘油致偏头痛大鼠发病机制中的作用探讨[D].中南大学硕士学位论文, 2009.