吗啡依赖及戒断大鼠中枢CCK-8基因表达的变化
发布时间：2019-05-07 04:33
【摘要】： 目的:药物依赖(drug dependence)是一种慢性、复发性的脑疾病,可引起大脑发生病理性变化。在我国滥用的毒品主要是阿片类物质,阿片类物质依赖包括身体依赖(表现为停药后的戒断症状)和精神依赖(表现为满足感和重新用药的渴求)。与阿片依赖及耐受相关的因素有内源性阿片肽、阿片受体及其受体后信号转导系统功能状态的适应性改变等。胆囊收缩素(cholecystokinin,CCK)最早发现于肠道并作为胃肠激素调节胆囊收缩和胰腺的生长分泌,后来的研究发现CCK广泛存在于中枢和周围神经系统,作为神经递质或调质发挥重要作用。CCK-8是目前已知的作用最强的内源性抗阿片肽,已有研究表明,CCK-8参与慢性吗啡依赖及戒断,但作用机制尚不清楚。本实验通过观察慢性吗啡依赖及戒断大鼠额叶皮质、尾壳核、海马CCK-8基因表达的变化,以探讨CCK-8在吗啡依赖及戒断反应中可能的作用。 方法:健康雄性Wistar大鼠36只,体重180～200 g,随机分组,每组12只,实验分为:生理盐水对照组(NS组)、吗啡依赖组(MOR组)、纳络酮催促戒断组(NAL组)。以剂量递增法(10mg·kg-1, 20mg·kg-1, 30mg·kg-1,40mg·kg-1, 50mg·kg-1)连续5天皮下注射吗啡建立吗啡依赖模型,以纳络酮5mg·kg-1单次腹腔注射建立吗啡依赖催促戒断模型,观察纳络酮催促戒断症状并评价造模是否成功;纳络酮戒断后1小时处死大鼠取额叶皮质、尾壳核、海马,每组取6只应用反转录聚合酶链反应(RT-PCR)检测CCK-8 mRNA含量;每组另外6只应用免疫组化(IHC)检测额叶皮质、尾壳核、海马CCK-8蛋白含量。用SPSS 11.5统计软件对实验数据进行分析,以P0.05为差异有显著性。 结果: 1.吗啡依赖动物模型的建立NS组和MOR组大鼠体重分别下降1.85±0.62g和2.63±0.8g,二者无显著差异,而NAL组大鼠体重下降(13.67±2.5g)较MOR组明显增加(P0.01);NS组和MOR组大鼠无一例发生跳跃反应,NAL组在注射纳络酮后的15min内跳跃次数达到33.7±3.99次,与MOR组相比明显增加(P0.01);注射纳洛酮后,NAL组大鼠均出现湿狗样震颤、不同程度的激惹、咬牙、流涎、腹泻以及扭体,戒断症状分值明显高于NS和MOR组(P0.01),其中NS组和MOR组大鼠评分均为0分,而NAL组大鼠的评分达到32.7±3.62。表明成功建立了吗啡身体依赖和纳络酮催促戒断动物模型。 2.吗啡依赖及戒断大鼠额叶皮质、尾壳核、海马CCK-8 mRNA含量的变化 MOR组大鼠尾壳核CCK-8 mRNA含量(0.609±0.030)较NS组(0.458±0.062)显著增高(P0.01),NAL组CCK-8 mRNA含量(0.437±0.059)较MOR组显著降低(P0.01),NAL组CCK-8 mRNA含量与NS组之间无明显差异(P0.05);额叶皮质、海马CCK-8 mRNA含量在各组之间无明显差异(P0.05)。 3.吗啡依赖及戒断大鼠额叶皮质、尾壳核、海马CCK-8蛋白含量的变化 MOR组大鼠尾壳核CCK-8蛋白含量(1.300±0.067)较NS组(0.956±0.058)显著增高(P0.01),NAL组CCK-8蛋白含量(0.956±0.084)较MOR组显著降低(P0.01),NAL组CCK-8蛋白含量与NS组之间无明显差异(P0.05);额叶皮质、海马CCK-8蛋白含量在各组之间无明显差异(P0.05)。 结论: 1.慢性吗啡依赖大鼠尾壳核CCK-8基因表达升高,纳络酮能够翻转此效应。 2.慢性吗啡依赖及纳络酮催促戒断大鼠,额叶皮质、海马CCK-8基因表达无变化。
[Abstract]:Objective: Drug dependence is a chronic and recurrent brain disease that can cause pathological changes in the brain. The drug abuse in our country is mainly opioids, and opioid dependence includes physical dependence (manifested as withdrawal symptoms after withdrawal) and mental dependence (manifested as a craving for satisfaction and remedication). The factors associated with opioid dependence and tolerance have endogenous opioid peptides, opioid receptors, and adaptive changes in the functional state of the signal transduction system after its receptor. Cholecystokinin (CCK) was first found in the intestinal tract and used as a gastrointestinal hormone to regulate the contraction of the gallbladder and the growth and secretion of the pancreas. CCK-8 is the most potent endogenous anti-opioid peptide currently known, and it has been shown that CCK-8 is involved in chronic morphine dependence and withdrawal, but the mechanism of action is not clear. The changes of the expression of CCK-8 gene in the frontal cortex, caudate nucleus and hippocampus of rats with chronic morphine dependence and withdrawal were observed to explore the possible role of CCK-8 in morphine dependence and withdrawal. Methods:36 healthy male Wistar rats were randomly divided into two groups: the normal saline control group (NS group), the morphine dependent group (MOR group) and the naloxone urging withdrawal group (NAL). The morphine dependence model was established by subcutaneous injection of morphine for 5 days in a dose-escalation method (10 mg 路 kg-1,20 mg 路 kg-1,30 mg 路 kg-1,40 mg 路 kg-1,50 mg 路 kg-1). The model was used to observe the effect of naloxone on the withdrawal symptoms and to evaluate the success of the model. One hour after the withdrawal of naloxone, the rats were sacrificed to take the cortex of the frontal lobe, the nucleus of the caudate shell and the hippocampus, and 6 of each group were used to detect the CCK-8 mRNA by reverse transcription polymerase chain reaction (RT-PCR). The content of the frontal cortex, the caudate nucleus and the CCK-8 protein in the frontal cortex, the caudate shell and the hippocampus were detected by immunohistochemistry (IHC) in each group. The data of the experimental data were analyzed by SPSS 11.5. The difference between the data and the experimental data was 0.05. The writing. Results:1. The weight of rats in NS group and MOR group decreased 1.85%, 0.62g and 2.63-0.8g, respectively. There was no significant difference between the two groups. In a case of a jump reaction, the number of hops in the NAL group was 33.7-3.99 times within 15 minutes after injection of naloxone, and compared with that of the MOR group (P0.01); after injection of naloxone, the rats of the nal group had a wet dog-like tremor, a different degree of agitation, a bite, a flow, The scores of salivation, diarrhea and torsion and withdrawal symptoms were significantly higher than that of NS and MOR (P0.01). The scores of rats in NS group and MOR group were 0, while the score of the rats in the NAL group was 3. 2.7 Figure 3.62. The successful establishment of morphine body-dependent and naloxone 2. Morphine-dependent and withdrawal rat frontal cortex, caudate nucleus, and hippocampus CC The content of CCK-8 mRNA in the rat tail shell of the modified MOR group (0.609-0.030) was significantly higher than that of the NS group (0.458-0.062) (P0.01), and the content of CCK-8 mRNA in the NAL group (0.437-0.059) was higher than that in the NS group (P 0.01). The levels of CCK-8 mRNA and CCK-8 mRNA in the frontal cortex and hippocampus in the frontal cortex and hippocampus were significantly lower than that in the NS group (P0.05). 3. There was no significant difference between morphine dependence and the frontal cortex of the withdrawal rats (P0.05). The content of CCK-8 (1.300-0.067) in the tail-shell nucleus and the content of the CCK-8 protein in the hippocampus was significantly higher than that in the NS group (0.956-0.058) (P0.01), and the content of the CCK-8 protein in the NAL group (0.956-0.067). There was no significant difference between the content of CCK-8 and the NS group (P <0.01), and there was no significant difference between the content of CCK-8 and the NS group (P0.05). white content There was no significant difference between the groups (P0.05). Conclusion:1. Chronic morphine dependence is large the expression of CCK-8 gene in the rat tail shell was increased, and the effect of naloxone could be reversed.2. Chronic morphine dependence and
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：D919
本文编号：2470776
[Abstract]:Objective: Drug dependence is a chronic and recurrent brain disease that can cause pathological changes in the brain. The drug abuse in our country is mainly opioids, and opioid dependence includes physical dependence (manifested as withdrawal symptoms after withdrawal) and mental dependence (manifested as a craving for satisfaction and remedication). The factors associated with opioid dependence and tolerance have endogenous opioid peptides, opioid receptors, and adaptive changes in the functional state of the signal transduction system after its receptor. Cholecystokinin (CCK) was first found in the intestinal tract and used as a gastrointestinal hormone to regulate the contraction of the gallbladder and the growth and secretion of the pancreas. CCK-8 is the most potent endogenous anti-opioid peptide currently known, and it has been shown that CCK-8 is involved in chronic morphine dependence and withdrawal, but the mechanism of action is not clear. The changes of the expression of CCK-8 gene in the frontal cortex, caudate nucleus and hippocampus of rats with chronic morphine dependence and withdrawal were observed to explore the possible role of CCK-8 in morphine dependence and withdrawal. Methods:36 healthy male Wistar rats were randomly divided into two groups: the normal saline control group (NS group), the morphine dependent group (MOR group) and the naloxone urging withdrawal group (NAL). The morphine dependence model was established by subcutaneous injection of morphine for 5 days in a dose-escalation method (10 mg 路 kg-1,20 mg 路 kg-1,30 mg 路 kg-1,40 mg 路 kg-1,50 mg 路 kg-1). The model was used to observe the effect of naloxone on the withdrawal symptoms and to evaluate the success of the model. One hour after the withdrawal of naloxone, the rats were sacrificed to take the cortex of the frontal lobe, the nucleus of the caudate shell and the hippocampus, and 6 of each group were used to detect the CCK-8 mRNA by reverse transcription polymerase chain reaction (RT-PCR). The content of the frontal cortex, the caudate nucleus and the CCK-8 protein in the frontal cortex, the caudate shell and the hippocampus were detected by immunohistochemistry (IHC) in each group. The data of the experimental data were analyzed by SPSS 11.5. The difference between the data and the experimental data was 0.05. The writing. Results:1. The weight of rats in NS group and MOR group decreased 1.85%, 0.62g and 2.63-0.8g, respectively. There was no significant difference between the two groups. In a case of a jump reaction, the number of hops in the NAL group was 33.7-3.99 times within 15 minutes after injection of naloxone, and compared with that of the MOR group (P0.01); after injection of naloxone, the rats of the nal group had a wet dog-like tremor, a different degree of agitation, a bite, a flow, The scores of salivation, diarrhea and torsion and withdrawal symptoms were significantly higher than that of NS and MOR (P0.01). The scores of rats in NS group and MOR group were 0, while the score of the rats in the NAL group was 3. 2.7 Figure 3.62. The successful establishment of morphine body-dependent and naloxone 2. Morphine-dependent and withdrawal rat frontal cortex, caudate nucleus, and hippocampus CC The content of CCK-8 mRNA in the rat tail shell of the modified MOR group (0.609-0.030) was significantly higher than that of the NS group (0.458-0.062) (P0.01), and the content of CCK-8 mRNA in the NAL group (0.437-0.059) was higher than that in the NS group (P 0.01). The levels of CCK-8 mRNA and CCK-8 mRNA in the frontal cortex and hippocampus in the frontal cortex and hippocampus were significantly lower than that in the NS group (P0.05). 3. There was no significant difference between morphine dependence and the frontal cortex of the withdrawal rats (P0.05). The content of CCK-8 (1.300-0.067) in the tail-shell nucleus and the content of the CCK-8 protein in the hippocampus was significantly higher than that in the NS group (0.956-0.058) (P0.01), and the content of the CCK-8 protein in the NAL group (0.956-0.067). There was no significant difference between the content of CCK-8 and the NS group (P <0.01), and there was no significant difference between the content of CCK-8 and the NS group (P0.05). white content There was no significant difference between the groups (P0.05). Conclusion:1. Chronic morphine dependence is large the expression of CCK-8 gene in the rat tail shell was increased, and the effect of naloxone could be reversed.2. Chronic morphine dependence and
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：D919
【参考文献】
相关期刊论文 前9条
1 闫玉仙;呼文亮;丛斌;马春玲;倪志宇;牛增强;余磊;;大鼠海马神经元中μ阿片受体、CCK受体的表达及慢性吗啡作用对其表达的影响[J];第四军医大学学报;2007年13期
2 谷建平,丛斌,刘东刚,谷振勇,毕海涛,朱桂云;吗啡依赖和戒断大鼠中枢cAMP和cGMP含量的变化[J];中国法医学杂志;2004年05期
3 洪仕君;李俊麟;李利华;瞿勇强;赵永和;;吗啡依赖鼠成瘾相关脑区腺苷酸环化酶的变化[J];法医学杂志;2006年04期
4 赵丽;丛斌;吴嫒嫒;李淑瑾;闫玉仙;姚玉霞;倪志宇;;吗啡对原代培养大鼠海马神经元CCK受体mRNA表达的影响[J];中国药理学通报;2006年04期
5 臧梦维,刘景生;NO-cGMP信息通路在阿片类药物耐受和依赖中的调节作用[J];中国药理学通报;1999年02期
6 胡晓东,李凤源;阿片类对钙通道的影响[J];中国药物依赖性杂志;1999年03期
7 谢燕,余争平,朱光绪,方强,江海洪;吗啡对培养海马神经元钙离子作用的机制研究[J];中国应用生理学杂志;2002年02期
8 李国海,黄明生,姜厚壁,俞俊洪;吗啡依赖大鼠脑内相关脑区CREB mRNA的表达[J];中国神经精神疾病杂志;2002年05期
9 方芳,汪青,曹清,刘景生;吗啡依赖小鼠脑组织cAMP系统的变化及腺苷酸环化酶磷酸化调节[J];中国医学科学院学报;2000年01期
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