中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2009年
11期
1008-1012
,共5页
闫玉仙%许顺江%丛斌%马春玲%余磊%牛增强%倪志宇%董玫
閆玉仙%許順江%叢斌%馬春玲%餘磊%牛增彊%倪誌宇%董玫
염옥선%허순강%총빈%마춘령%여뢰%우증강%예지우%동매
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%钙调素依赖性蛋白激酶2型
受體%膽囊收縮素B%嗎啡依賴%納洛酮%物質戒斷綜閤徵%海馬%神經元%鈣%鈣調蛋白%鈣%鈣調素依賴性蛋白激酶2型
수체%담낭수축소B%마배의뢰%납락동%물질계단종합정%해마%신경원%개%개조단백%개%개조소의뢰성단백격매2형
Receptor%cholecystokinin B%Morphine dependence%Naloxone%Substance withdrawal syndrome%Hippocampus%Neurons%Calcium%Calmodulin%Calcium-calmodulin-dependent protein kinase type 2
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)、钙凋蛋白(CaM)活性和钙调蛋白依赖性蛋白激酶Ⅱα(CaMKⅡα)表达的影响.方法 健康成年雄性Wistar大鼠45只,体重180～240 g,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1 h,腹腔注射纳洛酮5 mg/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1 h,相应靶核团注射10μg CR-2945,30 min后腹腔注射纳洛酮5 mg/kg;急性腹腔给药组(ACA组)于末次皮下注射吗啡后1 h,腹腔注射1 mg/kg CR-2945,30 min后腹腔注射纳洛酮5 mg/kg;慢性腹腔给药组(ACC组)给予吗啡的同时,腹腔注射1 mg/kg CR-2945,连续6 d,末次给药后30 min腹腔注射纳洛酮5 mg/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.
目的 探討膽囊收縮素-B(CCK-B)受體拮抗劑--CR-2945對嗎啡依賴大鼠納洛酮催促戒斷時海馬神經元內遊離Ca~(2+)濃度([Ca~(2+)]i)、鈣凋蛋白(CaM)活性和鈣調蛋白依賴性蛋白激酶Ⅱα(CaMKⅡα)錶達的影響.方法 健康成年雄性Wistar大鼠45隻,體重180～240 g,隨機分為9組(n=5):嗎啡依賴組(MD組)採用劑量遞增法建立嗎啡依賴大鼠模型;生理鹽水組(Ns組)以等容量生理鹽水替代嗎啡;納洛酮催促戒斷組(NPW組)于末次皮下註射嗎啡後1 h,腹腔註射納洛酮5 mg/kg;伏覈給藥組(NA組)、杏仁覈給藥組(A組)和側腦室給藥組(LCV組)于末次皮下註射嗎啡後1 h,相應靶覈糰註射10μg CR-2945,30 min後腹腔註射納洛酮5 mg/kg;急性腹腔給藥組(ACA組)于末次皮下註射嗎啡後1 h,腹腔註射1 mg/kg CR-2945,30 min後腹腔註射納洛酮5 mg/kg;慢性腹腔給藥組(ACC組)給予嗎啡的同時,腹腔註射1 mg/kg CR-2945,連續6 d,末次給藥後30 min腹腔註射納洛酮5 mg/kg;安慰劑組(P組)以生理鹽水替代CR-2945,其餘處理同LCV組.處理完畢後冰浴下分離海馬,採用流式細胞技術檢測海馬神經元內[Ca~(2+)]i和CaM活性,採用Western blot法測定CaMKⅡα錶達.結果 與NS組比較,MD組[Ca~(2+)]i和CaM活性升高,CaMKⅡα錶達上調(P<0.01);與MD組比較,NPW組[Ca~(2+)]i和CaM活性降低,CaMKⅡα錶達下調(P<0.01);與NPW組比較,ACA組、ACC組和LCV組[Ca~(2+)]i和CaM活性升高,CaMKⅡα錶達上調(P<0.01),NA組、A組和P組上述指標差異無統計學意義(P>0.05).結論 CCK-B受體拮抗劑通過升高海馬神經元內[Ca~(2+)]i、CaM活性,上調CaMKⅡα錶達,從而抑製嗎啡依賴大鼠納洛酮催促戒斷反應.
목적 탐토담낭수축소-B(CCK-B)수체길항제--CR-2945대마배의뢰대서납락동최촉계단시해마신경원내유리Ca~(2+)농도([Ca~(2+)]i)、개조단백(CaM)활성화개조단백의뢰성단백격매Ⅱα(CaMKⅡα)표체적영향.방법 건강성년웅성Wistar대서45지,체중180～240 g,수궤분위9조(n=5):마배의뢰조(MD조)채용제량체증법건립마배의뢰대서모형;생리염수조(Ns조)이등용량생리염수체대마배;납락동최촉계단조(NPW조)우말차피하주사마배후1 h,복강주사납락동5 mg/kg;복핵급약조(NA조)、행인핵급약조(A조)화측뇌실급약조(LCV조)우말차피하주사마배후1 h,상응파핵단주사10μg CR-2945,30 min후복강주사납락동5 mg/kg;급성복강급약조(ACA조)우말차피하주사마배후1 h,복강주사1 mg/kg CR-2945,30 min후복강주사납락동5 mg/kg;만성복강급약조(ACC조)급여마배적동시,복강주사1 mg/kg CR-2945,련속6 d,말차급약후30 min복강주사납락동5 mg/kg;안위제조(P조)이생리염수체대CR-2945,기여처리동LCV조.처리완필후빙욕하분리해마,채용류식세포기술검측해마신경원내[Ca~(2+)]i화CaM활성,채용Western blot법측정CaMKⅡα표체.결과 여NS조비교,MD조[Ca~(2+)]i화CaM활성승고,CaMKⅡα표체상조(P<0.01);여MD조비교,NPW조[Ca~(2+)]i화CaM활성강저,CaMKⅡα표체하조(P<0.01);여NPW조비교,ACA조、ACC조화LCV조[Ca~(2+)]i화CaM활성승고,CaMKⅡα표체상조(P<0.01),NA조、A조화P조상술지표차이무통계학의의(P>0.05).결론 CCK-B수체길항제통과승고해마신경원내[Ca~(2+)]i、CaM활성,상조CaMKⅡα표체,종이억제마배의뢰대서납락동최촉계단반응.
Objective To investigate the effect of CCK-B receptor antagonist CR-2945 on cytoplasmic free calcium concentration ([Ca~(2+)]i), calmodulin (CaM) activity and calmodulin-dependent protein kinase Ⅱα(CaMK Ⅱα) 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 AC A) , 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. In 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 10μg 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. CaMK Ⅱα expression was determined by Western blot. Results [Ca~(2+) ]i, and CaM activity were significantly increased and CaMK Ⅱα expression was up-regulated in group MD compared with group NS (P < 0.01). [Ca~(2+)]i, and CaM activity were significantly decreased and CaMK Ⅱ a expression was down-regulated in group NPW compared with group MD ( P<0.01) . [Ca~(2+)]i, and CaM activity were significantly increased and CaMK Ⅱ a expression was up-regulated in group ACA, ACC and LCV compared with group NPW ( P < 0.01 ), but no significant change in the indices mentioned above was found in group NA, A and P ( P > 0.05) . Conclusion CCK-B receptor antagonist CR-2945 can inhibit naloxone-precipitated withdrawal responses through increasing [Ca~(2+)]i and CaM activity and up-regulating CaMK Ⅱα expression in hippocampal neurons in morphine-dependent rats.
