瑞芬太尼诱发切口痛大鼠痛觉过敏时脊髓NR2B与CaMKⅡα的关系  

Relationship between NR2B and CaMKⅡα in spinal cord during remifentanil-induced hyperalgesia in a rat model of incisional pain

作　　者：徐如彬 王春艳[2] 李依泽[2] 张麟临 贾韡 赵亓[2] 郭素倩 王国林[2] Xu Rubin;Wang Chunyan;Li Yize;Zhang Linlin;Jia Wei;Zhao Qi;Guo Suqian;Wang Guolin(Tianjin Medical University First Center Clinical College, Department of Anesthesiology, Tianjin First Center Hospital, Tianjin 300192, China;Department of Anesthesiology, Tianjin Medical University General Hospital Tianjin Research Institute of Anesthesiology, Tianjin 300052, China)

机构地区：[1]天津医科大学一中心临床学院、天津市第一中心医院麻醉科,300192 [2]天津医科大学总医院麻醉科、天津市麻醉学研究所,300052

出　　处：《中华麻醉学杂志》2018年第10期1209-1213,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81600962,81371245,81500961,81400908).

摘　　要：目的评价瑞芬太尼诱发切口痛大鼠痛觉过敏时脊髓含2B亚基的NMDA受体(NR2B)与钙/钙调素依赖性蛋白激酶Ⅱα(CaMKⅡα)的关系。方法取尾静脉置管及鞘内置管成功的雄性SD大鼠40只,体重260~280g,2~3月龄,采用随机数字表法分为4组(n=10):对照组(C组)鞘内注射生理盐水0.1ml,10min后经尾静脉输注生理盐水0.1ml·kg^-1·min^-1,持续60min;瑞芬太尼+切口痛组(RI组)鞘内注射生理盐水0.1ml,10min后尾静脉输注瑞芬太尼1.0μg·kg^-1·min^-1,持续60min,输注开始即刻建立切口痛模型;NR2B拮抗剂Ro25-6981组(Ro组)鞘内注射Ro25-6981(0.1ml)10μg,10min后尾静脉输注生理盐水0.1ml·kg^-1·min^-1,持续60min;瑞芬太尼+切口痛+Ro25-6981组(RI+Ro组)鞘内注射Ro25-6981(0.1ml)10μg,10min后尾静脉输注瑞芬太尼1.0μg·kg^-1·min^-1,持续60min,输注开始即刻制备切口痛模型。于静脉输注生理盐水或瑞芬太尼前24h、输注完毕后2、6、24和48h(T0-4)时测定机械缩足反应阈(MWT)和热缩足潜伏期(TWL)。最后一次行为学测试后处死大鼠,取L4-6脊髓节段,采用Westernblot法测定脊髓总蛋白和膜蛋白NR2B(tNR2B和mNR2B)及总蛋白CaMKⅡα(tCaMKⅡα)和磷酸化CaMKⅡα(pCaMKⅡα)的表达水平,计算mNR2B/tNR2B比值及pCaMKⅡα/tCaMKⅡα比值。结果与C组比较,RI组MWT降低,TWL缩短,脊髓tNR2B、mNR2B,tCaMKⅡα及pCaMKⅡα表达上调,mNR2B/tNR2B比值、pCaMKⅡα/tCaMKⅡα比值升高(P<0.05或0.01)。与RI组比较,RI+Ro组MWT升高,TWL延长,脊髓tNR2B、mNR2B和tCaMKⅡα及pCaMKⅡα表达下调,mNR2B/tNR2B比值、pCaMKⅡα/tCaMKⅡα比值降低(P<0.05或0.01)。结论瑞芬太尼诱发切口痛大鼠痛觉过敏时脊髓NR2B功能增强可激活CaMKⅡα,该过程可能参与了瑞芬太尼诱发术后痛觉过敏形成的机制。Objective To evaluate the relationship between NR2B subunit-containing N-methyl-D-aspartate (NMDA) receptors (NR2B receptors) and Ca2+ /calmodulin-dependent protein kinase Ⅱ α (CaMKⅡα) in the spinal cord during remifentanil-induced hyperalgesia in a rat model of incisional pain (IP). Methods Forty male Sprague-Dawley rats in which intrathecal and caudal catheters were successfully placed, weighing 260-280 g, aged 2-3 months, were divided into 4 groups (n=10 each) using a random number table method: control group (group C), remifentanil plus IP group (group RI), NR2B antagonist Ro 25-6981 group (group Ro) and remifentanil plus IP plus Ro 25-6981 group (group RI+ Ro). In group C, normal saline 0.1 ml was intrathecally injected, and 10 min later normal saline was infused for 60 min via the tail vein at a rate of 0.1 ml·kg^-1·min^-1.In group RI, normal saline 0.1 ml was intrathecally injected, and 10 min later remifentanil was infused for 60 min via the tail vein at a rate of 1.0 μg· kg^-1·min^-1, and IP was established immediately after onset of remifentanil infusion.In group Ro, Ro 25-6981 (0.1 ml) 10 μg was intrathecally injected, and 10 min later normal saline was infused for 60 min via the tail vein at a rate of 0.1 ml·kg^-1·min^-1. In group RI+ Ro, Ro 25-6981 (0.1 ml) 10 μg was intrathecally injected, and 10 min later remifentanil was infused for 60 min via the tail vein at a rate of 1.0 μg· kg^-1·min^-1, and IP was established immediately after onset of remifentanil infusion.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before intravenously infusing normal saline or remifentanil and at 2, 6, 24 and 48 h after the end of infusion (T0-4). The rats were sacrificed after the last behavioral test, and the L4-6 segment of the spinal cord was removed for determination of the expression of NR2B in total and membrane protein (tNR2B and mNR2B) and expression of CaMKⅡα in total protein(tCaMKⅡα) and phosphorylated CaMKⅡα (pCaMKⅡα)

关 键 词：哌啶类 痛觉过敏 受体 N-甲基-D-天冬氨酸 钙-钙调素依赖性蛋白激酶类 

分 类 号：R614[医药卫生—麻醉学]