NLRP3炎性小体在慢性偏头痛小鼠三叉神经脊束尾核的表达及作用  

The role of NLRP3 inflammasome in trigeminal nucleus caudalis of mice with chronic migraine

作　　者：贺维[1] 龙婷 张艺馨 秦光成[1] 陈力学[1] 周冀英[2] HE Wei;LONG Ting;ZHANG Yi-Xin;QIN Guang-Cheng;CHEN Li-Xue;ZHOU Ji-Ying(Experimental Research Center,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Chongqing Key Laboratory of Neurology,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院实验研究中心,重庆400016 [2]重庆市神经病学重点实验室,重庆400016

出　　处：《中国疼痛医学杂志》2020年第7期490-495,共6页Chinese Journal of Pain Medicine

基　　金：国家自然科学基金(81671092);重庆市渝中区科技计划项目(201701106)。

摘　　要：目的:观察炎性小体NLRP3(Nod-like receptor protein 3)在慢性偏头痛小鼠三叉神经脊束尾核(trigeminal nucleus caudalis,TNC)中的表达变化,并探讨其在慢性偏头痛中的作用。方法:C57BL6/J雄性小鼠随机分为5组,分别为生理盐水对照组(n=16),模型组(n=16),模型+溶剂组(n=8),模型+NLRP3抑制剂5 mg/kg组(n=8),模型+NLRP3抑制剂10 mg/kg组(n=8)。采用腹腔隔日重复注射硝酸甘油(nitroglycerin,NTG)建立慢性偏头痛小鼠模型。使用von Frey test测定小鼠眶周和后足机械痛阈值。Western blot检测NLRP3及神经元活化标志蛋白p-ERK的表达变化,免疫荧光检测NLRP3在TNC的定位情况。结果:反复腹腔注射NTG后,小鼠眶周和后足的机械痛阈显著降低(P<0.05),而TNC部位的NLRP3和p-ERK蛋白表达显著增加(P<0.05)。使用NLRP3的小分子抑制剂MCC950可以显著提高小鼠眶周和足底机械痛阈值,降低p-ERK的表达。荧光染色显示NLRP3与小胶质细胞共定位。结论:本研究表明TNC部位NLRP3炎性小体上调,可能通过影响神经元活化状态参与慢性偏头痛的病理生理过程。Objective:To investigate the role of NLRP3 inflammasome within trigeminal mucleus caudalis(TNC)in the pathophysiological process of chronic migraine(CM)mice.Methods:Male C57BL6/J mice were randomly divided into 5 groups:saline group(n=16),CM group(n=16),CM+PBS group(n=8),CM+MCC950(5 mg/kg)group(n=8)and CM+MCC950(10 mg/kg)group(n=8).Nitroglycerin(NTG)was repeatedly intraperitoneally injected(i.p.)in awake mice to establish CM model.Mechanical pain thresholds of periorbital region and hind paw were detected by von Frey test.Western blotting was performed to detect the expression of NLRP3 and p-ERK protein in TNC.Localization of NLRP3 in TNC was detected by immunofluorescence methods.Results:Repeated injection of NTG decreased pain thresholds in both periorbital region and mechanical withdrawal threshold(MWT)of the hind paw(P<0.05),and up-regulated the expression of NLRP3 and p-ERK in TNC(P<0.05).Furthermore,inhibition of NLRP3 by MCC950 relieved the allodynia and reduced the expression of p-ERK.Immunofluorescence staining revealed that NLRP3 was expressed in microglia in TNC.Conclusion:The up-regulation of NLRP3 inflammasome in TNC may be involved in the pathophysiological process of allodynia in CM by affecting the activation status of neurons.

关 键 词：NLRP3炎性小体 慢性偏头痛 小胶质细胞 三叉神经脊束尾核 

分 类 号：R747.2[医药卫生—神经病学与精神病学]