PICK1在瑞芬太尼痛觉过敏大鼠脊髓含GluR1及GluR2亚基的AMPA受体转运中的作用  

Role of PICK1 in trafficking of GIuR1-and GluR2-containing AMPA receptors in spinal cord in a rat model of remifentanil-induced hyperalgesia

作　　者：王志芬[1] 王国林[1] 敖吉莹 丁玲[1] 李楠[1] 汤晓红[1] 张麟临 舒瑞辰 元元[1] 

机构地区：[1]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》2015年第8期927-931,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81300960,81371245）;天津市应用基础及前沿技术研究计划（14JCQNJC12800）;天津市卫生局科技基金（2013KZ124）

摘　　要：目的 评价蛋白激酶Cα相互作用蛋白1 （PICK1）在瑞芬太尼痛觉过敏大鼠脊髓含GluR1及GluR2亚基的AMPA受体转运中的作用.方法 SPF级雄性SD大鼠32只,体重240 ～260 g,42 ～ 49日龄,鞘内置管成功后,采用随机数字表法分为4组（n=8）：对照组（C组）、生理盐水＋瑞芬太尼组（NS＋R组）、PICK1反义核苷酸＋生理盐水组（AS＋NS组）和P1CK1反义核苷酸＋瑞芬太尼组（AS＋R组）.C组、NS＋R组鞘内注射生理盐水10μl,AS＋NS组、AS＋R组鞘内注射硫代修饰的PICK1反义核苷酸10 μg/10μl,1次/d,连续4d.鞘内注射结束后,NS＋R组和AS＋R组静脉输注瑞芬太尼1.2 μg·kg-1·min-160 min,C组和AS＋NS组静脉输注等容量生理盐水60 min.于静脉输注生理盐水或瑞芬太尼前24 h、输注结束后2、6、24和48 h（T0-4）时测定机械缩足反应阈（MWT）和热缩足潜伏期（TWL）.最后一次痛阈测定结束后处死大鼠,采用Western blot法测定脊髓细胞膜及细胞浆含GluR1及GluR2亚基的AMPA受体的表达水平.计算细胞膜与细胞浆蛋白表达水平的比值（m/c比值）和细胞膜含GluR1及GluR2亚基的AMPA受体的表达水平的比值（mGluR 1/mGluR2比值）.结果 与C组比较,NS＋R组和AS＋R组T1-4时TWL缩短,MWT降低,细胞膜含GluR1亚基的AMPA受体表达上调,其m/c比值升高,细胞膜含GluR2亚基的AMPA受体表达下调,其m/c比值降低,mGluR 1/mGluR2比值升高（P＜0.05）.与NS＋R组比较,AS＋R组T1-4时TWL延长,MWT升高,细胞膜含GluR2亚基的AMPA受体表达上调,其m/c比值升高,mGluR 1/mGluR2比值降低（P＜0.05）,其余指标差异无统计学意义（P＞0.05）.结论 PICK1可促进脊髓含GluR2亚基的AMPA受体内化,而对含GluR1亚基的AMPA受体转运无影响,该作用可能参与了瑞芬太尼诱发大鼠痛觉过敏形成的机制.Objective To evaluate the role of protein interacting with Cα kinase 1 （PICK1 in trafficking of GIuR1-and GluR2-containing AMPA receptors in the spinal cord in a rat model of remifentanilinduced hyperalgesia.Methods Thirty-two male Sprague-Dawley rats, weighing 240-260 g, aged 42-49 days, in which IT catheters were successfully placed, were randomly divided into 4 groups （n =8 each） using a random number table： control group （group C） , normal saline＋remifentanil group （group NS＋ R） , PICK1 antisense oligonucleotide ＋ normal saline group （group AS ＋ NS） , and PICKI antisense oligonucleotide＋remifentanil group （group AS＋R）.Normal saline 10 μl was injected intrathecally in C and NS＋R groups, and thiolmodificated PICK1 antisense oligonucleotide 10 μg/10 μl was injected intrathecally in AS＋NS and AS＋R groups, once a day for 4 consecutive days.After the end of IT injection, remifentanil was infused intravenously at 1.2 μg · kg-1 · min-1 for 60 min in NS＋R and AS＋R groups, while the equal volume of normal saline was given instead for 60 min in C and AS＋ NS groups.The mechanical paw withdrawal threshold （MWT） and thermal paw withdrawal latency （TWL） were measured at 24 h before normal saline or remifentanil infusion, and at 2, 6, 24, and 48 h after the end of normal saline or remifentanil infusion.The rats were sacrificed after the last measurement of pain threshold.The lumbar segment L4-6 of the spinal cord was removed for detection of the expression of GluR1 and GluR2-containing AMPA receptors in cell membrane and in cytoplasm by Western blot.The ratio of protein expression in cell membrane to that in cytoplasm （m/c ratio） was calculated.The ratio of GluRl-containing AMPA receptor expression to GluR2-containing AMPA receptor expression in cell membrane was calculated （mGluR1/ mGluR2）.Results Compared with group C, the TWL was significantly shortened, and the MWT was decreased at T1-4, the expression of GluR1-containing AMPA receptors in cell membra

关 键 词：载体蛋白质类 哌啶类 痛觉过敏 受体 AMPA 

分 类 号：R614[医药卫生—麻醉学]