杏仁核MCP-1/CCR2通路在大鼠神经病理性痛中的作用  

Role of monocyte chemoattractant factor/chemokine C-C receptor 2pathway in amygdala in neuropathic pain in rats

作　　者：杨冯睿 易汉 彭良玉[2] 胡啸玲[2] 郭曲练[1] Yang Fengrui;Yi Han;Peng Liangyu;Hu Xiaoling;Guo Qulian(Department of Anesthesiology,Xiangya Hospital of Central South University,Changsha 410008,China;Department of Anesthesiology,First Affiliated Hospital of University of South China,Hengyang 421001,China)

机构地区：[1]中南大学湘雅医院麻醉科,长沙市410008 [2]南华大学附属第一医院麻醉科,衡阳市421001

出　　处：《中华麻醉学杂志》2018年第7期850-854,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81300971);湖南省自然科学基金(2016JJ4079)。

摘　　要：目的 评价杏仁核单核细胞趋化因子-1(MCP-1)∕细胞表面趋化因子受体2( CCR2)通路在大鼠神经病理性痛中的作用.方法 清洁级健康雄性SD大鼠,体重200~260 g,2月龄,采用结扎左侧L5,6脊神经法制备神经病理性痛模型.实验Ⅰ 取大鼠32只,采用随机数字表法分为2组:对照组(C组,n=8)和神经病理性痛组(NP组,n=24). NP组于神经病理性痛模型制备后第7、14和21 天,C组相应于模型制备前,处死后取杏仁核组织,分别采用实时定量 PCR和免疫组化法检测MCP-1、CCR2的mRNA表达和阳性细胞数.实验Ⅱ 取大鼠32只,采用随机数字表法分为4组(n=8):C组、NP组、MCP-1组及CCR2特异性拮抗剂RS102895组( RS组). MCP-1组和RS组于神经病理性痛模型制备后第3、6、13和20天双侧杏仁核分别注射MCP-1 (每侧50 pmol)和 RS102895 (每侧100 pmol).于神经病理性痛模型制备前( T0)、制备后第4、7、14、21 天( T1-4)时测定热缩足潜伏期(TWL)和机械缩足反应阈(MWT);于T4时处死大鼠,取L5脊髓组织,采用ELISA法测定IL-1β、IL-6和TNF-α含量.结果 实验Ⅰ 与C组比较,NP组杏仁核MCP-1和CCR2的mRNA表达上调,阳性细胞数增多(P<0.05).实验Ⅱ 与C组比较,其余3组T1-4时MWT降低,TWL缩短,脊髓IL-1β、IL-6和TNF-α含量升高(P<0.05).与NP组比较,MCP-1组T1时MWT降低,TWL缩短,脊髓IL-1β、IL-6和TNF-α含量升高;RS组T1-4时MWT升高,TWL延长,脊髓IL-1β、IL-6和TNF-α含量降低(P<0.05或0. 01).结论 杏仁核MCP-1∕CCR2通路功能增强可能参与了大鼠神经病理性痛的病理生理过程.Objective To evaluate the role of monocyte chemotactic factor-1 (MCP-1)∕chemokine C-C receptor 2 ( CCR2) in amygdala in neuropathic pain ( NP) in rats. Methods Clean-grade healthy male Sprague-Dawley rats, weighing 200-260 g, aged 2 months, in which NP model was established by ligating the left L5,6spinal nerve, were used in this study. The experiment was performed in two parts. Ex-periment Ⅰ Thirty-two rats were divided into 2 groups using a random number table method: control group (C group, n=8) and NP group (n=24). Rats were sacrificed at 7, 14 and 21 days after establis-hing NP model in group NP or at the corresponding time points before establishing NP model in group C, and the amygdala was removed for determination of the expression of MCP-1 and CCR2 mRNA and positive cell count using quantitative real-time polymerase chain reaction and immunohistochemistry. Experiment ⅡThirty-two rats were divided into 4 groups ( n=8 each) using a random number table method: control group (C group), NP group, MCP-1 group and specific CCR2 antagonist RS102895 group (RS group). MCP-1 (50 pmol for each side) or RS102895 (100 pmol for each side) was injected into the bilateral a-mygdala at days 3, 6, 13 and 20 after establishing NP model in MCP-1 and RS groups, respectively. The thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at days 4, 7, 14 and 21 after establishing NP model (T1-4). Rats were sacrificed at T4and the L5segment of the spinal cord was removed for determination of interleukin-1beta (IL-1β), IL-6 and tumor necrosis fac-tor-alpha ( TNF-α) contents by enzyme-linked immunosorbent assay. Results Experiment Ⅰ Compared with group C, the expression of MCP-1 and CCR2 mRNA in amygdala was significantly up-regulated, and the number of MCP-1 and CCR2 positive cells was increased in group NP ( P<0. 05). Experiment ⅡCompared with group C, the MWT was significantly decreased and TWL was shortened at T1-4, and the contents of IL-1β, IL-6 and TNF-α were increased in

关 键 词：神经痛 趋化因子CCL2 受体 CCR2 杏仁核 

分 类 号：R741[医药卫生—神经病学与精神病学]