复方戒毒肽对吗啡成瘾小鼠脑内N-甲基-D-天门冬氨酸受体1和一氧化氮合酶表达水平的影响

Effect of compound detoxification peptide on the levels of N-methyl-D-aspartate receptor 1 and nitric oxide synthase in the brain of mice with morphine addiction

目的:观察复方戒毒肽对吗啡成瘾小鼠脑内N-甲基-D-天门冬氨酸受体1和一氧化氮合酶水平的影响。方法:实验于2003-03/2004-05在郑州大学基础医学院生物化学与分子生物学教研室动物室和郑州大学志愿戒毒研究中心第一实验室完成。复方戒毒肽由脑肽配制而成熏主要含有酸性肽,牛神经肽FF等小分子神经肽。80只健康的昆明种小鼠,雌雄各半,8～10周龄,体质量(20±2)g。抽签随机分为8组,每组10只。正常对照组:不做任何处理。成瘾模型建立:腹腔注射吗啡,1次/d熏100mg/kg,共11d。成瘾模型组:成瘾模型建立后,不做任何处理。生理盐水组:成瘾模型建立后,用生理盐水1mL/次灌胃治疗15d。自然戒断组:成瘾模型建立后,正常喂养15d,期间不做任何处理;高、中、低剂量的复方戒毒肽治疗组:成瘾模型建立后,给成瘾小鼠按80mg/kg,40mg/kg,20mg/kg的复方戒毒肽进行灌胃治疗,神经肽溶液的实际用量为1mL熏0.5mL熏0.25mL,时间为15d。单独使用复方戒毒肽组:仅给正常小鼠按80mg/kg剂量的复方戒毒肽灌胃15d。实验结束后取材,通过免疫组化和Biosens数字成像系统分析仪测定海马内N-甲基-D-天门冬氨酸受体1和一氧化氮合酶水平。结果:80只小鼠均进入结果分析。各组N-甲基-D-天门冬氨酸受体1和一氧化氮合酶的水平的比较:与正常对照组相比,成瘾模型组两者水平均增高(134.93±6.83,117.24±3.91;116.26±8.64,96.81±6.16,P<0.05);与成瘾模型组、自然戒断组、生理盐水组、40mg/kg和20mg/kg复方戒毒肽组相比,80mg/kg复方戒毒肽组两者水平均明显降低(134.93±6.83,117.24±3.91;133.10±8.97,115.82±5.61;134.92±8.51,116.55±5.10;120.02±7.49,106.03±8.99;120.02±7.49,106.03±8.99;128.59±9.26,112.59±2.93;130.50±2.50,113.22±5.14,P<0.05)。结论:80mg/kg的复方戒毒肽能够降低吗啡成瘾小鼠脑内N-甲基-D-天门冬氨酸受体1和一氧化氮合酶的水平。

AIM： To study the effect of compound detoxification peptide on the levels of N-methyl-D-aspartate receptor 1 （NMDAR1） and nitric oxide synthase （NOS） in mice with morphine addiction. METHODS： The experiment was conducted in the Animal Test Room of Department of Biochemistry and Molecular Biology of the Basic Medical College as well as the First Laboratory for Research Center of Refraining Drugs of Zhengzhou University from March 2003 to May 2004. Compound detoxification peptide was prepared by brain peptide, mainly including acidic poptide bovine neuropeptide and some other micromolecular neuropoptides. Eighty healthy Kunming mice （half female and half male） with an age ranged 8-10 weeks and the body mass of （20±2） g were selected and randomly divided into 8 groups with 10 mice in each group. Mice in the normal control group were not interfered. Establishment of animal models of morphine addiction： No treatment was conducted on the models. Normal saline group： Gastric perfusion of normal saline was given to models of morphine addiction at 1 mL each time for totally 15 days. Natural refraining group： Mice were normally fed for 15 days after establishment of morphine addiction models without any other treatments. High, middle, low dose compound detoxification peptide treatment groups： Mice with morphine addiction were given gastric perfusion of 80 mg/kg, 40 mg/kg, 20 mg/kg of compound refraining peptide at the actual neuropeptide dose of 1 mL, 0.5 mL, 0.25 mL respectively for totally 15 days. Simple compound refraining poptide group： mice were only given gastric perfusion of compound refraining peptide 80 mg/kg for 15 days. Materials were obtained after the experiment, and the levels of NMDAR1 and NOS were measured histochemical method and Biosens Digital Analyzer. RESULTS： A total of 80 mice were involved in the analysis of results. Comparison in level of NMDAR1 and NOS among each group： compared with the normal control group, those in the morphine addiction model group were increased （134.93±6.83, 117.24±3.91; 116.26±8.64, 96.81±6.16, P 〈 0.05）, while those were significantly decreased in the 80 mg/kg compound refraining peptide group than those in the morphine addition group, natural refraining group, normal saline group, 40 mg/kg, 20 mg/kg compound refraining poptide group 134.93±6.83, 117.24±3.91; 133.10±8.97, 115.82±5.61; 134.92±8.51, 116.55±5.10; 120.02±7.49, 106.03±8.99; 120.02±7.49, 106.02±8.99; 128.59±926, 112.59±2.93; 130.50±2.50, 113.22±5.14, P 〈 0.05）. CONCLUSION： The levels of NUDAR1 and NOS in brain of mice with morphine addiction can be decreased by 80 mg/kg of compound refraining poptide.