| 题名 | 吗啡影响免疫系统细胞大麻素受体的表达及机体免疫功能 |
| 作者 | 张庆余 |
| 学位类别 | 硕士 |
| 答辩日期 | 2011-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 曹毅 |
| 关键词 | 吗啡滥用 大麻素受体 阿片受体 免疫机能失调 Morphine abuse Cannabinoid receptor Opioid receptor Immune dysfunction |
| 其他题名 | Morphine affects the cannabinoid receptor expression in immune cells and immune function |
| 学位专业 | 细胞生物学 |
| 中文摘要 | 吗啡滥用（morphine abuse）导致机体免疫功能受损是影响毒品吸食人员生存质量的问题之一。阿片受体与大麻素受体是两类与吗啡成瘾相关的受体分子，它们不但分布于神经系统神经元细胞，亦分布于免疫细胞膜表面。分布于免疫细胞表达的吗啡相关受体介导并影响了免疫细胞的免疫反应性，包括免疫细胞病原吞噬、抗原递呈、免疫细胞迁移、免疫球蛋白合成及细胞因子释放等过程。阿片与大麻素受体介导的信号转导的异常导致免疫系统细胞功能异常，引起免疫系统机能紊乱。本论文研究了吗啡滥用与免疫系统细胞中大麻素受体表达之间的关系。论文分为三部分。 （一） 为研究吗啡吸食人员免疫系统免疫机能及免疫细胞中吗啡相关受体的变化情况，我们检测了吗啡吸食且美沙酮维持治疗人员（methadone maintenance treatment and physical dependence group）和有吗啡吸食但已戒断人员（morphine withdrawal and physical independence group）的血浆免疫球蛋白（IgG和IgM）和免疫炎症因子IL-4和IFN-γ在外周血单核细胞中mRNA的表达。同时也检测了其外周血单核细胞（periphery blood mononuclear cell，PBMC）中与吗啡相关的阿片受体与大麻素受体及与这两类受体介导的信号通路密切相关的转录因子(CREB、c-fos和STAT6)的mRNA表达。吸毒人员的所有指标与正常健康无吸毒经历人员的对应指标进行统计学比较，实验结果显示免疫球蛋白IgG和IgM相对于正常健康无吸毒经历人员相比均升高，且升高后二者含量的比例与升高前无差异；免疫炎症因子IL-4在吸毒人员血浆当中升高，IFN-γ含量在吸毒人员当中降低；三种阿片受体表达在吸毒人员与正常健康人之间无差异；在吸毒人员中大麻素受体1（CB1r）与大麻素受体2（CB2r）相对于正常健康无吸毒经历人员中mRNA表达上调，且大麻素受体1上升程度相对大麻素受体2更明显。CREB蛋白是大麻素受体介导的信号通路（Gi蛋白偶联受体信号通路）中一个重要细胞转录因子，我们实验结果表明CREB（cAMP respond element binding protein）在吸食吗啡人员PBMC细胞中的表达相对于健康无吸毒人员降低；另一个与大麻受体信号通路相关的转录因子c-fos（即刻早期基因）在接受美沙酮维持治疗人员中相对于吗啡吸食但已戒断人员于和正常健康无吸毒经历人员(healthy group)表达上升，而在未接受治疗的两组人员间无差异。与c-fos 基因表达变化情况类似，调控大麻素受体基因表达的细胞转录因子STAT6（signal transducer and activator of transcription 6）在吸食吗啡且美沙酮维持治疗组中相对于吗啡吸食但已戒断和健康无吸毒经历组升高，而在未接受治疗的两组人员间无差异。我们结果表明在吗啡滥用人员中免疫系统重要的免疫球蛋白及免疫炎症因子的水平相对于健康无吸毒经历的人员均有不同程度的改变，说明吗啡滥用人员免疫系统功能异常，这可能与CREB、c-fos、STAT6、大麻素受体的变化相关。 （二） 为证明吗啡是否可以在直接作用于细胞，改变细胞大麻素受体基因的表达。我们使用不同浓度吗啡刺激体外培养的神经、免疫系统的细胞，一段时间后收集并检测这两种细胞大麻素受体的蛋白和mRNA表达水平。发现吗啡刺激前，神经来源和免疫来源的细胞都有不同程度及不同种类的大麻素受体的表达。吗啡刺激后，大麻素受体mRNA和蛋白质表达均有改变，且这种改变有时间依赖性和剂量依赖性。这表明吗啡可以直接作用于神经或者免疫系统细胞，直接调节大麻素受体基因的表达。 （三） 为应用树鼩动物模型研究毒品成瘾对神经/免疫系统的影响提供基础数据,我们对大麻素及阿片受体在正常猕猴和树鼩神经系统和免疫系统的表达进行初步确定。采集正常猕猴和树鼩新鲜组织(皮质、小脑、脑干、海马、脊髓、脾脏),检测大麻素与阿片受体mRNA在猕猴和树鼩各组织中的表达情况，我们实验结果表明在树鼩中，重要脑区（皮层、小脑、脑干、海马）、脊髓及代表性免疫器官脾脏大麻素受体2和mu型阿片受体均有表达，大麻素受体1在除脾脏外其他组织也均有表达。两种阿片受体（delta,kappa ）在各组织中未检测到表达。在猕猴组织中，大麻素受体1和mu型阿片受体在所有检测的组织中都有表达；kappa型阿片受体在皮质、小脑、脑干、脊髓中有表达；大麻素受体2仅在脾脏中表达；delta型阿片受体仅在海马中检测到有表达。我们证实了与吗啡作用相关的阿片受体和大麻素受体在树鼩的各个重要脑区、脊髓和脾脏的表达模式与猕猴和人都较接近，这表明树鼩作为毒品成瘾的模型动物有利用潜能。 |
| 英文摘要 | Morphine abuse impaired immune system is one of the problems which influence the morphine abusers’ quality of life. Opioid and cannabinoid receptors are related with morphine addiction and they are not only distributed in the neurons but also on the surface of immune cells. The receptors are located on the surface of immune cells and regulate the cell functions including the pathogen phagocytosis, antigen presentation, immune cell migration, immunoglobulin synthesis and cytokines release. Abnormal alteration of opioid and cannabinoid receptors induces immune disorder. Here,we investigated the relationship between morphine abuse and the expressions of cannabinoid receptors in the immune cells the study included the following three parts. (Ⅰ)To study the immune function and the alteration of morphine addiction related receptors in themethadone maintenance treatment ；CREB protein play an important role on the GPCR (G-protein coupled receptor) family signal pathway, it is a mediator of signal transduction of opioid and cannabinoid receptor. CREB expression is down-regulated in the PBMC cells of morphine abusers. Immediately early gene c-fos which is also an important transcription factor is involved in activation of cannabinoid and opioid receptor. Expression of c-fos was up-regulated in methadone maintenance treatment group compared with the morphine withdrawal group and the healthy group. However, there is no difference between the morphine withdrawal group and the healthy group. Similarly, the STAT6 gene expression was up-regulated in the methadone maintenance treatment group compared with the morphine withdrawal group and the healthy group. There is no statistically significant difference between the morphine withdrawal group and the healthy group. These results show that immumoglobin and inflammatory cytokines levels were changed in morphine abusers.the alteration of immune function may be involved in STAT,CREB,c-fos and cannabinoid receptors.people and morphine withdrawal people. We tested the levels of immunoglobin (IgG and IgM) and the proinflammatory cytokines in the serum of methadone maintenance treatment people and morphine withdrawal people .We also tested the mRNA expression of morphine related receptor and important transcription factor involved in the signal pathway mediated by cannabinoid receptor in the peripheral blood monocular cell (PBMC) of the maintenance treatment people and the morphine withdrawal people. Our results suggest that both of IgG and IgM contents were increased but the ratio of IgG and IgM still have no difference between the morphine abuse people (methadone maintenance treatment people and the morphine withdrawal people) and the healthy people; Proinlfammatory IL-4 level is increased and IFN-γ level is decreased in the morphine abusers compared with the healthy people; the expressions of the three opioid receptors were not affected; However, expression of the two kinds of cannabinoid receptors were up-regulated, and cannabinoid receptor 1(CB1r) expression level was severely increased compared with cannabinoid receptor 2(CB2r). |
| 语种 | 中文 |
| 公开日期 | 2013-04-23 |
| 内容类型 | 学位论文 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7379]  |
| 专题 | 昆明动物研究所_分子病理学 |
| 推荐引用方式 GB/T 7714 | 张庆余. 吗啡影响免疫系统细胞大麻素受体的表达及机体免疫功能[D]. 北京. 中国科学院研究生院. 2011. |
| 个性服务 |
| 查看访问统计 |
| 相关权益政策 |
| 暂无数据 |
| 收藏/分享 |
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。
修改评论