JNK在神经病理性痛大鼠中脑导水管周围灰质星形胶质细胞活化中的作用  

Role of c-Jun N-terminal kinase in activation of astrocytes in midbrain periaqueductal gray of rats with neuropathic pain

作　　者：倪华栋[1] 安康 姚明[1] 黎亮[2] 黄冰[1] 费勇[1] 王云贡 王寒琪 朱春燕 陈文宇[3] Ni Huadong;An Kang;Yao Ming;Li Liang;Huang Bing;Fei Yong;Wang Yungong;Wang Hanqi;Zhu Chunyan;Chen Wenyu(Department of Anesthesiology and Pain Medical Center, First Affiliated Hospital of Jiaxing University, Jiaxing 314001, China;Department of General Surgery, First Affiliated Hospital of Jiaxing University, Jiaxing 314001, Chin;Department of Pneumology, First Affiliated Hospital of Jiaxing University, Jiaxing 314001, China;Department of Anesthesiology, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Chin)

机构地区：[1]嘉兴学院附属第一医院麻醉与疼痛医学中心,314001 [2]嘉兴学院附属第一医院肝胆外科,314001 [3]嘉兴学院附属第一医院呼吸科,314001 [4]温州医科大学附属第二医院麻醉科,325000

出　　处：《中华麻醉学杂志》2017年第12期1450-1453,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81341035）;浙江省医药卫生科技计划一般项目（2015KYA217）;浙江省卫生高层次创新人才培养工程项目（2012-RC-22）;浙江省浙北区域麻醉专病中心基金（2015-24）;浙江省自然科学基金（LY16H090016）;嘉兴市科技计划项目（2017AY33008）

摘　　要：目的评价c-Jun氨基末端激酶（JNK）在神经病理性痛大鼠中脑导水管周围灰质（PAG）星形胶质细胞活化中的作用。方法清洁级雄性SD大鼠72只，9周龄，体重160～200 g，采用随机数字表法分为4组：正常对照组（C组，n＝8）、神经病理性痛组（NP组，n＝40）、二甲基亚砜溶剂对照组（DS组，n＝12）和JNK抑制剂SP600125组（SP组，n＝12）。采用慢性坐骨神经缩窄性损伤（CCI）法制备大鼠神经病理性痛模型。于CCI后14 d时，SP组PAG内注射10 nmol JNK抑制剂SP600125 0.5 μl，DS组注射10%二甲基亚砜0.5 μl。C组取8只大鼠，NP组于CCI前、CCI后3、7、14、21 d时取8只大鼠，DS组和SP组取8只大鼠分别于CCI前、CCI后14 d给药前、给药后30、45、60、75和90 min时测定机械痛阈。C组机械痛阈测定结束后处死，取PAG组织，采用Western blot法测定PAG区磷酸化JNK（p-JNK）和胶质纤维酸性蛋白（GFAP）的表达；NP组于各时点痛阈测定结束后处死，测定PAG区p-JNK表达；DS组和SP组取4只大鼠于给药后45 min时机械痛阈测定结束后处死，测定PAG区GFAP的表达。结果与C组比较，NP组CCI后各时点机械痛阈降低，CCI后7～21 d时p-JNK表达上调（P〈0.01）；与DS组比较，SP组给药后30 min时机械痛阈升高，给药后45 min时GFAP表达下调（P〈0.01）；与给药前比较，SP组给药后30～75 min时机械痛阈升高（P〈0.01）。结论神经病理性痛大鼠PAG星形胶质细胞活化机制与JNK激活有关。Objective To evaluate the role of c-Jun N-terminal kinase（JNK）in activation of astrocytes in midbrain periaqueductal gray（PAG）of rats with neuropathic pain.Methods A total of 72 pathogen-free male Sprague-Dawley rats, aged 9 weeks, weighing 160-200 g, were divided into 4 groups using a random number table： control group（group C, n=8）, neuropathic pain group（group NP, n=40）, dimethyl sulfoxide control group（group DS, n=12）and JNK inhibitor SP600125 group（group SP, n=12）. Neuropathic pain was produced by chronic constriction injury（CCI）. At 14 days after CCI, 10 nmol JNK inhibitor SP600125 0.5 μl was intraperitoneally injected into the PAG in group SP, and 10% dimethyl sulfoxide 0.5 μl was given instead in group DS.Eight rats were selected in group C, before CCI and at 3, 7, 14 and 21 days after CCI in group NP, and in DS and SP groups, and the mechanical pain threshold was measured before CCI, before administration on 14 days after CCI and at 30, 45, 60, 75 and 90 min after administration.The rats in group C were sacrificed after the end of measurement of the mechanical pain threshold, and brains were removed for determination of phosphorylated JNK（p-JNK）and glial fibrillary acidic protein expression（by Western blot）in PAG region.The rats in group NP were sacrificed after the end of measurement of the mechanical pain threshold at each time point, and brains were removed for detection of p-JNK expression in PAG region.Four rats in DS and SP groups were sacrificed after the last measurement of the mechanical pain threshold at 45 min after administration, and brains were removed for determination of glial fibrillary acidic protein expression in PAG region.Results Compared with group C, the mechanical pain threshold was significantly decreased at each time point after CCI, and the expression of p-JNK was up-regulated at 7-21 days after CCI in group NP（P〈0.01）. Compared with group DS, the mechanical pain threshold was significantly increased at 30 min after administration,

关 键 词：星形细胞 JNK丝裂原活化蛋白激酶类 脑导水管 水管周灰质 神经痛 

分 类 号：R402[医药卫生—临床医学]