脊髓胶质细胞p38有丝分裂原活化蛋白激酶在大鼠持续性术后痛形成中的作用：与Toll样受体4的关系  

Role of p38 mitogen-activated protein kinase in spinal cord in development of persistent postoperative pain in rats： the relationship with Toll-like receptor 4

作　　者：胡兴国[1] 阳红艳[2,3,4] 文锟 张云翔[1] 曾因明[2,3] 

机构地区：[1]湖南省桃源县人民医院麻醉危重医学科,415700 [2]徐州医学院江苏省麻醉与镇痛应用技术重点实验室 [3]江苏省麻醉学重点实验室 [4]浙江大学邵逸夫医院麻醉科

出　　处：《中华麻醉学杂志》2014年第5期574-577,共4页Chinese Journal of Anesthesiology

基　　金：湖南省科技计划一般项目资助课题（2009SK3018）;江苏省教育厅高校省级重点实验室开放课题（KJS1106）

摘　　要：目的 评价脊髓胶质细胞p38有丝分裂原活化蛋白激酶（p38MAPK）在大鼠持续性术后痛形成中的作用及其与Toll样受体4（TLR4）的关系.方法 鞘内置管成功的大鼠120只,体重200～250 g,2月龄,采用随机数字表法,将其分为5组（n=24）：假手术组（S组）、皮肤/肌肉切口和牵拉组（SMIR组）、SMIR＋二甲基亚砜（DMSO）组（DMSO组）、SMIR＋ p38MAPK抑制剂SB203580组（SB203580组）和SMIR＋ TLR4小干扰RNA（TLR4siRNA）组（TLR4siRNA组）.制备SMIR诱发持续性术后痛模型,DMSO组和SB203580组于术前30 min和术后1-12 d分别鞘内注射2％DMSO 10μl和SB203580（5 μg）,TLR4siRNA组于术前1d和术后1-12 d鞘内给予TLR4siRNA（2 μg）,1次/d.分别于术前1d及术后1、3、7、12、22 d时测定机械缩足反应阈（MWT）,各时点MWT测定结束后处死4只大鼠,取L4-6节段脊髓,采用Western blot法测定磷酸化p38MAPK（p-p38MAPK）表达.结果 与S组比较,SMIR组和DMSO组术后MWT降低,脊髓p-p38MAPK表达上调（P＜0.05）;与SMIR组比较,SB203580组和TLR4siRNA组术后MWT升高,脊髓p-p38MAPK表达下调（P＜0.05）,DMSO组各时点MWT和脊髓p-p38MAPK表达差异无统计学意义（P＞0.05）.结论 脊髓胶质细胞TLR4通过激活p38MAPK参与了大鼠持续性术后痛的形成.Objective To evaluate the role of p38 mitogen-activated protein kinase （p38MAPK） in the spinal cord in the development of persistent postoperative pain evoked by skin/muscle incision and retraction （SMIR） and the relationship with Toll-like receptor 4 （TLR4）.Methods One hundred and twenty male SpragueDawley rats,weighing 200-250 g,aged 2 months,in which intrathecal catheters were successfully implanted,were randomly divided into 5 groups （n =24 each） using a random number table：sham operation group （group S）,SMIR group,SMIR ＋ dimethyl sulfoxide （DMSO） group （group DMSO）,SMIR ＋ p38MAPK inhibitor SB203580 group （group SB203580） and SMIR ＋ TLR4 small interference RNA （siRNA） group （group TLR4siRNA）.The rats were anesthetized with intraperitoneal chloral hydrate 400 mg/kg.The skin and superficial muscle of the medial thigh were incised and a small pair of retractors inserted.This tissue was retracted for 1 h causing potential stretch of the saphenous nerve.2% DMSO 10 μl and SB203580 5 μg were injected intrathecally at 30 min before operation and 1-12 days after operation in DMSO and SB203580 groups,respectively.TLR4siRNA 2 μg was administered intrathecally at 1 day before operation and 1-12 days after operation once a day in group TLR4siRNA.Mechanical paw withdrawal threshold to von Frey filament stimulation （MWT） was measured at 1 day before operation and 1,3,7,12 and 22 days after operation.Four rats in each group were sacrificed after measurement of MWT at each time point,and the L4-6 segments of the spinal cord were obtained for detection of the expression of phosphorylated p38MAPK （p-p38MAPK） by Western blot analysis.Results Compared with group S,MWT was significantly decreased after operation,and the expression of p-p38MAPK was up-regulated after operation in SMIR and DMSO groups.Compared with group SMIR,MWT was significantly increased after operation,and the expression of p-p38MAPK was down-regulated after operation in SB203580 and TLR4siRNA groups,and

关 键 词：TOLL样受体4 神经胶质 疼痛 手术后 P38丝裂原活化蛋白激酶类 脊髓 

分 类 号：R619[医药卫生—外科学]