CCK—B受体拮抗剂对吗啡依赖大鼠纳洛酮催促戒断时海马神经元[Ca^2＋]i、CaM活性和CaMKⅡα表达的影响  

Effect of CCK-B receptor antagonist on [ Ca^2＋ ] i, CaM activity and CaMKⅡα expression in hippocampal neurons during naloxone-peecipitated withdrawal in morphine-dependent rats

作　　者：闫玉仙[1] 许顺江[2] 丛斌[1] 马春玲[1] 余磊[1] 牛增强[1] 倪志宇[1] 董玫[1] 

机构地区：[1]河北医科大学法医学系河北省法医学重点实验室,石家庄市050017 [2]河北医科大学第一医院中心实验室

出　　处：《中华麻醉学杂志》2009年第11期1008-1012,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金资助项目（30672355）;河北省自然科学基金资助项目（C2007000826）

摘　　要：目的探讨胆囊收缩素-B（CCK—B）受体拮抗剂——CR-2945对吗啡依赖大鼠纳洛酮催促戒断时海马神经元内游离Ca^2＋浓度（[Ca^2＋]i）、钙调蛋白（CaM）活性和钙调蛋白依赖性蛋白激酶Ⅱ12（CaMKⅡα）表达的影响。方法健康成年雄性Wistar大鼠45只，体重180—240g，随机分为9组（n=5）：吗啡依赖组（MD组）采用剂量递增法建立吗啡依赖大鼠模型；生理盐水组（NS组）以等容量生理盐水替代吗啡；纳洛酮催促戒断组（NPW组）于末次皮下注射吗啡后1h，腹腔注射纳洛酮5mg／kg；伏核给药组（NA组）、杏仁核给药组（A组）和侧脑室给药组（LCV组）于末次皮下注射吗啡后1h，相应靶核团注射10ug CR-2945，30min后腹腔注射纳洛酮5mg／kg；急性腹腔给药组（ACA组）于末次皮下注射吗啡后1h，腹腔注射1mg／kgCR-2945，30min后腹腔注射纳洛酮5mg／kg；慢性腹腔给药组（acc组）给予吗啡的同时，腹腔注射1mg／kgCR-2945，连续6d，末次给药后30min腹腔注射纳洛酮5mg／kg；安慰剂组（P组）以生理盐水替代CR-2945，其余处理同LCV组。处理完毕后冰浴下分离海马，采用流式细胞技术检测海马神经元内[Ca^2＋]i和CaM活性，采用Western blot法测定CaMKⅡα表达。结果与Ns组比较，MD组[Ca^2＋]i和CaM活性升高，CaMKⅡα表达上调（P〈0．01）；与MD组比较，NPW组[Ca^2＋]i和CaM活性降低，CaMKⅡα表达下调（P〈0．01）；与NPW组比较，ACA组、ACC组和LCV组[Ca^2＋]i和CaM活性升高，CaMKⅡα表达上调（P〈0．01），NA组、A组和P组上述指标差异无统计学意义（P〉0．05）。结论CCK—B受体拮抗剂通过升高海马神经元内[Ca^2＋]i、CaM活性，上调CaMKⅡα表达，从而抑制吗啡依赖大鼠纳洛酮催促戒断反应。Objective To investigate the effect of CCK-B receptor antagonist CR-2945 on cytoplasmic free calcium concentration （[Ca^2＋] i）, ealmodulin （CAM） activity and calmodulin-dependent protein kinase Ⅱα （CaMKⅡα） expression in hippocampal neurons during naloxone-precipitated withdrawal in morphine-dependent rats. Methods Forty-five adult male Wistar rats weighing 180-240 g were randomly divided into 9 groups （ n= 5 each）： morphine dependence group （group MD）, normal saline group （group NS）, naloxone-precipitated withdrawal group （group NPW）, nucleus accumbens group （group NA）, amygdale group （group A）, lateral cerebral ventricle group （group LCV）, abdominal cavity （acute） group （group ACA）, abdominal cavity （chronic） group （group ACC） and placebo group （group P）. Morphine dependence was induced by increasing doses of subcutaneous injection of morphine for 5 d and subcutaneous injection of morphine 50 mg/kg at 8：00 on 6th day. hi group NS, equal volume of normal saline was injected instead of morphine. In group NPW, intraperitoneal nalxone 5 mg/kg was injected 1 h after the last subcutaneous injection of morphine. In group NA, A and LCV, CR-2945 10 ug was injected into nucleus accumbens, amygdale and lateral ventricle 1 h after the last subcutaneous injection of morphine, and then intraperitoneal naloxone 5 mg/kg was injected 30 min later. In group ACA, CR- 2945 1 mg/kg was injected 1 h after the last subcutaneous injection of morphine, and then intraperitoneal naloxone 5 mg/kg was injected 30 min later. In group ACC, CR-2945 1 mg/kg was injected simultaneously with morphine injection for 6 d, and intraperitoneal naloxone 5 mg/kg was injected 30 min after the last administration. In group P, normal saline was used instead of CR-2945, and the other procedures were the same as those in group LCV. Rats were killed and hippocampus was isolated. [ Ca^2＋] i and CaM activity were measured by flow cytometry. CaMKII c~ expression was determined by Western b

关 键 词：受体 胆囊收缩素B 吗啡依赖 纳洛酮 物质戒断综合征 海马 神经元 钙 钙调蛋白 钙-钙凋素依赖性蛋白激酶2型 

分 类 号：R749.64[医药卫生—神经病学与精神病学]