吗啡依赖、戒断及丁丙诺啡的替代治疗对大鼠垂体POMC基因表达的影响

目的··:结合戒断体征的观察从分子水平研究吗啡依赖和戒断机理及丁丙诺啡对吗啡戒断的影响。方法··:观察吗啡戒断及经丁丙诺啡治疗大鼠的戒断体征 ;利用核酸分子杂交技术研究其垂体阿黑皮源(POMC)基因表达的变化。结果··:丁丙诺啡能有效减轻或消除戒断体征,吗啡依赖及戒断时大鼠垂体POMC -mRNA含量下降,丁丙诺啡对其影响不大。结论··:吗啡依赖和戒断抑制大鼠垂体POMC基因表达,丁丙诺啡能纠正吗啡躯体依赖,但从基因水平看丁丙诺啡可能也具有依赖潜能     

咪唑啉受体及其对阿片药理作用的调节机制

用稳定转染咪唑啉1型受体(I1R)的细胞(CHO-I1),对I1R信号转导途径进行了初步研究。首次直接证实I1R的信号转导途径与活化PC-PLC继而产生DAG,其后引起丝裂原激活蛋白激酶(MAPK)酶促级联反应过程有关。采用共同稳定表达μ阿片受体(MOR)和I1R的CHO细胞表达系统(CHO-μ/I1细胞),对I1R在受体后水平抑制吗啡依赖的可能分子机制进行了研究。首次提供了胍丁胺通过作用于I1R而抑制吗啡依赖的直接实验证据,其分子机制可能主要是胍丁胺激活I1R抑制吗啡慢性处理时cAMP通路和Ca2+信号通路代偿性适应而抑制吗啡依赖的形成。     

慢性吗啡依赖大鼠脑内G_(i2)蛋白的改变

目的研究慢性吗啡依赖大鼠腹侧背盖区(Ventraltegmentalarea,VTA)、伏隔核(Nucleusaccumbens,NAc)、前额皮质(Prefrontalcortex,PC)、海马(Hippocampus)及去甲肾上腺能神经中枢蓝斑(Locuscoeruleus,LC)五个脑区Gi2蛋白的改变,探讨慢性吗啡依赖的可能机制。方法18只SD大鼠随机分为三组:慢性吗啡依赖组、戒断组和空白对照组。慢性吗啡依赖组和戒断组腹腔注射吗啡,建立吗啡成瘾模型,戒断组腹腔注射纳络酮(5mg/kg)30min后断头处死各组大鼠,取出脑组织,进行冰冻切片。用免疫组化技术检测NAc、PC、LC、VTA和Hippocampus五个脑区相对Gi2蛋白水平。结果慢性吗啡成瘾组和戒断组与空白对照组相比,NAc区Gi2蛋白水平明显降低(P<0.01),LC区Gi2蛋白水平却明显升高(P<0.01),其它脑区未发现明显变化。结论慢性吗啡成瘾可引起大鼠脑内Gi2蛋白水平发生改变,但各脑区Gi2蛋白水平的改变并不相同。Gi2蛋白水平的改变可能是吗啡耐受和依赖潜在的分子机制。     

吗啡对吗啡依赖大鼠腹侧苍白球神经元放电的影响

目的 :研究吗啡对吗啡依赖大鼠腹侧苍白球的电生理影响 ,探讨其在成瘾机制中的作用。方法 :通过单细胞细胞外记录吗啡依赖大鼠腹侧苍白球的电信号 ,观察吗啡对放电的影响。结果 :吗啡依赖大鼠腹侧苍白球神经元放电频率给药前为 3 76Hz±s 1 18Hz,注射吗啡后为 13 2 2Hz±s 2 37Hz;并且纳洛酮可以逆转这种兴奋效应。结论 :吗啡显著兴奋吗啡依赖大鼠腹侧苍白球神经元 ,腹侧苍白球在药物强化过程中起着重要作用。     

青藤碱对吗啡依赖小鼠脊髓与小脑中HO2、sGCα1和sGCα2基因mRNA表达的影响

目的评价青藤碱治疗前后吗啡依赖与戒断小鼠脊髓和小脑中HO2、sGCα1和sGCα2基因mRNA水平的变化,探讨青藤碱作用于吗啡依赖小鼠的分子机制。方法采用吗啡剂量递增法,建立小鼠吗啡依赖模型,用青藤碱(40mg.kg-1,ip)治疗吗啡依赖小鼠后,再用纳洛酮激发急性戒断症状,半定量RT-PCR测脊髓和小脑HO2、sGCα1和sGCα2基因的mRNA水平变化。结果慢性吗啡用药使小鼠脊髓与小脑中HO2和sGCα1基因的mRNA水平发生异常上调,sGCα2基因的mRNA水平没有异常上调。单独使用青藤碱,没有引起小鼠脊髓与小脑中这些指标异常升高。青藤碱治疗后,小鼠吗啡依赖与戒断时在脊髓与小脑中异常上调的HO2和sGCα1基因的mRNA水平下降到接近对照组水平。结论慢性吗啡用药使HO2和sGCα1基因的mRNA水平发生不同程度的异常上调,这种上调在青藤碱治疗后可以恢复到接近正常水平,这可能是青藤碱对吗啡依赖小鼠治疗作用的分子机制之一。     

吗啡依赖相关的酶学研究进展

吗啡依赖形成的机制是一个复杂的过程,涉及特定脑区突触超微结构变化、神经递质、酶学等,并且相互之间有密切的联系.近年国内外学者从多层次、多角度进行了研究,试图从根本上阐明吗啡依赖形成的机理.对吗啡依赖酶学方面的研究近年取得了较大进展,目前研究较多的吗啡依赖相关酶体系有蛋白激酶类、一氧化氮合酶、超氧化物歧化酶和腺苷酸环化酶等.现将吗啡依赖相关上述酶学体系的研究进展综述如下.     

吗啡在基因水平对脑嘌呤核苷酸代谢的影响

目的:通过研究吗啡对大鼠脑组织嘌呤核苷酸代谢的影响,寻找吗啡依赖相关的调控与效应基因。方法:利用核酸分子杂交测定吗啡依赖与戒断大鼠丘脑及颞叶脑组织次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)mRNA的含量变化。结果:核酸分子杂交密度扫描结果显示吗啡依赖、急性及慢性戒断时大鼠丘脑及颞叶脑组织HGPRTmRNA均明显低于对照组(P<0.05,P<0.01,P<0.001)。结论:吗啡能明显抑制丘脑及颞叶脑组织HGPRT的基因表达,提示吗啡可能通过调控脑核苷酸代谢的关键酶,影响脑核苷酸及核酸代谢。     

吗啡依赖与毒蕈碱型乙酰胆碱受体的适应

毒蕈碱型乙酰胆碱(muscarinic acetylcholine)能神经在阿片依赖中的作用可以追溯到30年代,当时认为吗啡可以抑制迷走神经.到60年代认为吗啡镇痛耐受与毒蕈碱乙酰胆碱能神经的慢性适应有关;70年代有作者认为吗啡戒断时毒蕈碱能神经兴奋[1].本文系统地介绍M受体介导吗啡镇痛耐受、吗啡戒断反应和吗啡精神依赖的作用机制.     

吗啡蛋白质组的研究概况

蛋白质组的研究已以成为人类基因计划完成后揭示复杂生物分子交互作用规律,是深入研究各种生化过程,推动医学向纵深发展的有力手段.从1975年逐步发展起来的双向电泳、生物质谱技术、电喷射离子化技术(ESI)和基质辅助激光解吸附离子化技术(MALDI)等,已提供了敏感而且明确的多肽和蛋白质及其氨基酸序列的鉴定方法.本文专门关注吗啡药物依赖的特定蛋白质组,即"吗啡蛋白质组(morphinome)"[1],简要介绍其研究状况及研究模型.     

蛋白质组学方法在研究阿片类物质依赖机理中的应用

阿片类物质成瘾（依赖）的机理十分复杂。大脑中有关组织中蛋白质表达改变可能与阿片类物质依赖产生密切相关。海马是一个调节学习记忆功能的重要脑组织，近年来研究表明它在阿片等毒品成瘾机理中也有重要作用。研究海马组织蛋白表达改变与阿片类物质成瘾的关系，对认识阿片类物质成瘾的机理和发现药物作用的靶标的重要意义。应用蛋白质组学方法，我们观察到慢性吗啡处理能引起小鼠海马中与能量代谢密切相关的呼吸链中的NADH脱氢酶的辅基Fe-s蛋白，三羧酸循环中丙酮酸脱氢酶复合物中二氢硫辛酰转乙酰基酶和糖酵解关键酶乳酸脱氢酶表达显著地降低。荧光实时定量PCR（RT-PCR）分析表明编码这三个蛋白质表达的mRNA含量相应地降低。吗啡慢性处理小鼠海马组织中的ATP水平显著地低于正常小鼠海马组织中ATP水平，进一步证实慢性吗啡依赖小鼠海马中三个与能量代谢相关的酶表达降低。     

中枢神经系统疾病及其药物与蛋白质组学研究进展

阐述中枢神经系统疾病及其药物蛋白质组学研究的最新进展。蛋白质组学是后基因组时代的一门重要学科 ,是从整体水平对蛋白质进行综合分析 ,目前已广泛应用于临床和生物医学各个领域。蛋白质组学研究有助于阐明CNS疾病发生、发展、转归的网络机制 ,寻找疾病特异性蛋白质 ,针对疾病靶点定向合成药物 ,构建分子药理模型 ,高通量地筛选和评价药物的效应及毒副作用。可以预见 ,蛋白质组学将为CNS疾病的诊断、监测和药物研制起到不可估量的作用     

An Integrated Quantitative Proteomics and Systems Biology Approach to Explore Synaptic Protein Profile Changes During Morphine Exposure

Morphine is a classic analgesic for the treatment of chronic pain. However, its repeated use is known to produce tolerance, physical dependence, and addiction; these properties limit its long-term therapeutic use and this has led to a quest for therapeutics without these unwanted side effects. Understanding the molecular changes in response to long-term use of morphine is likely to aid in the development of novel therapeutics for the treatment of pain. Studies examining the effects of chronic morphine administration have reported alterations in gene expression, synapse morphology, and synaptic transmission implying changes in synaptic protein profile. To fully understand the changes in protein profiles, proteomic techniques have been used. Studies using two-dimensional gel electrophoresis of various brain regions combined with mass spectrometry have found alterations in the levels of a number of proteins. However, neither the changes in brain regions relevant to morphine effects nor changes in the abundance of synaptic proteins have been clearly delineated. Recent studies employing subcellular fractionation to isolate the striatal synapse, combined with quantitative proteomics and graph theory-inspired network analyses, have begun to quantify morphine-regulated changes in synaptic proteins and facilitate the generation of networks that could serve as targets for the development of novel therapeutics for the treatment of chronic pain. Thus, an integrated quantitative proteomics and systems biology approach can be useful to identify novel targets for the treatment of pain and other disorders of the brain.     

阿片类药物对海马神经递质释放调控的分子机制

阿片类药物的滥用,可导致脑内神经元的长时程适应,从而产生严重的身体依赖和心理依赖。越来越多的证据表明,阿片类药物所引起的这种适应性变化会影响突触前神经递质的释放,并改变包括谷氨酸能神经系统在内的神经元的功能。我们的研究表明,在海马这个富含谷氨酸能神经元的脑区,阿片类药物如吗啡的长期使用,会引起胞外谷氨酸水平的持续改变。进一步的研究表明,该脑区的SNARE蛋白复合物发生了适应性的改变。SNARE蛋白家族是重要的调节神经可塑性的调节分子,在神经递质释放过程中起重要作用。在吗啡长期处理过程中,SNAP-25的色氨酸187位点的磷酸化明显下降,SNARE复合物的形成受到抑制。我们的结果揭示了阿片类药物可通过调控突触前直接参与递质释放的分子来影响递质本身释放这一分子机制。     

Inhibitory effects of ginseng total saponin on up-regulation of cAMP pathway induced by repeated administration of morphine

We have reported that ginseng total saponin (GTS) inhibited the development of physical and psychological dependence on morphine. However, the possible molecular mechanisms of GTS are unclear. Therefore, this study was undertaken to understand the possible molecular mechanism of GTS on the inhibitory effects of morphine-induced dependence. It has been reported that the up-regulated cAMP pathway in the LC of the mouse brain after repeated administration of morphine contributes to the feature of withdrawals. GTS inhibited up-regulation of cAMP pathway in the LC after repeated administration of morphine in this experiment. GTS inhibited cAMP levels and protein expression of protein kinase A (PKA). In addition, GTS inhibited the increase of cAMP response element binding protein (CREB) phosphorylation. Therefore, we conclude that the inhibitory effects of GTS on morphine-induced dependence might be mediated by the inhibition of cAMP pathway.     

分子生物学技术在中药领域的应用及其对中药新药研究的影响

分子生物学技术在中药领域已得到广泛应用,并对中药新药研究产生了深刻的影响.该文介绍了 PCR, RFLP, PCR- RFLP, RAPD等技术和方法在中药的分子鉴定与中药质量控制、中药进化与系统学及中药新品种选育中的应用,以及基因干预和蛋白质组学在中药作用机理研究与新药开发研究方面的应用和影响.     

大麻二酚在毒品依赖中的干预作用及其机制的研究进展

毒品滥用是当今世界严重的公共卫生和社会问题。由于其具有精神依赖性强、复吸率高和神经毒性大的特点,给家庭和社会带来严重的负面影响。然而针对毒品依赖的有效治疗手段仍然是当今医疗卫生事业的难点。大麻二酚是植物大麻中提取的非毒性成分,已在治疗多种中枢神经系统疾病(如帕金森病、癫痫、阿尔茨海默病等)方面具有潜在价值,受到人们广泛关注。最近研究表明,大麻二酚对吗啡、可卡因、甲基苯丙胺、苯丙胺,酒精等药物诱导的精神依赖具有良好的干预作用,但具体的机制仍然不清楚。该文对大麻二酚在干预毒品依赖中的作用及其相关机制进行综述,旨在进一步研究大麻二酚在治疗毒品依赖中的潜在价值和相关机制,发现新的防治毒品依赖的机制靶点提供重要参考。     

不同分型钙通道阻滞剂在吗啡成瘾中的作用

长期应用吗啡可产生成瘾,Ca2+参与镇痛和成瘾过程的形成。近年来研究发现不同分型钙通道阻滞剂对吗啡的镇痛、耐受和依赖等药理作用具有不同的影响,为进一步阐明吗啡成瘾机制和戒毒药物的研究提供了新的启示。     

Tumor vasculature directed drug targeting: applying new technologies and knowledge to the development of clinically relevant therapies

Recognition of the dependence of solid tumor growth on the formation of new blood vessels has ignited an enormous research effort aimed at the development of new therapeutic strategies for cancer. Besides direct application of drugs inhibiting endothelial cell function during angiogenesis, tumor vasculature directed drug-targeting strategies have been investigated for this purpose. In animal models of disease, proof of principle regarding the potential of selective interference with tumor blood flow as a powerful tumor therapy has been generated to its full extent. The challenge for the coming years will be to develop these strategies into clinically applicable ones. New insights into the molecular mechanisms prevailing in the endothelium during angiogenesis and into the mechanism(s) of action of drugs with anti-angiogenic activities, as well as new techniques to identify useful tumor endothelium specific target epitopes have in recent years been exploited to meet this challenge. This review summarizes vasculature directed therapeutic strategies proven to be successful in pre-clinical models and new (drug targeting) technologies enabling the development of more effective therapeutics for the treatment of cancer.     

The ongoing evolution of proteomics in malignancy

The complementary fields of genomics and proteomics offer insights into the molecular mechanisms of diseases. While genomics seeks to define our genetic substrate, proteomics explores the structure and function of proteins, which are the end effectors of our genes. Proteomics has been revolutionized in the past decade by the application of techniques such as protein arrays, two-dimensional gel electrophoresis, and mass spectrometry. These techniques have tremendous potential for biomarker development, target validation, diagnosis, prognosis, and optimization of treatment in medical care, especially in the field of clinical oncology. We will discuss innovations in proteomic technologies and highlight their prospective applications to patient care.