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神经元nAChR阻滞模型与抗衰老食品评价

Models Based on the Blockade on Neuronal Nicotinic Acetylcholine Receptors and Its Applications in Evaluation of Anti-Aging Food

【作者】 刘洁生

【导师】 彭志英;

【作者基本信息】 华南理工大学 , 食品工程, 1998, 博士

【摘要】 神经元烟碱乙酰胆碱受体(Nicotinic acetylcholine receptors,nAChRs)在动物学习记忆智能活动及人类某些认知退行性疾患中具有重要作用,近年来受到广泛的关注,是神经科学研究的前沿课题。 近年来中枢nAChR研究取得了突破性进展主要是依赖于K-神经毒素(Kappa-Neurotoxin K-NT)系列探针及其它探针的应用。由于K-NT探针来源受限制,至今有关中枢nAChR的功能研究的报道仍甚少。 本文在分离纯化K-NT的基础上,采用α-银环蛇毒素(α-Bungarotoxin,α-BGT)和K-NT制备了果蝇头部粗制膜受体模型和K-NT所致学习记忆障碍模型,并对一些抗衰益智食品进行了评价,取得了如下主要成果: 1.果蝇头部粗制膜受体及药理学特点 采用125I-α-BGT标记果蝇头部粗制膜受体,经Scatchard作图求得KD为5.0mmol/L,Bmax为350fmol/mg.α-BGT和K-NT对125I标记的α-BGT与果蝇头部nAChR结合具有拮抗作用,两者IC50和Ki值仅相差5倍,而在脊椎动物,K-BGT对电鳐电器官nAChRs的亲和力不足α-BGT的1/300,这表明果蝇头部nAChR是一兼性受体,具有神经-肌肉结点和神经元nAChR的特点。 2.果蝇头部nAChR的随龄变化及麦饭石的抗衰老作用评价 果蝇头部nAChR随龄变化,平衡常数KD值在1-30龄果蝇基本不变,而Bmax值则随增龄而减少,一日龄为400fmol/mg,而30日龄降低至260fmol/mg,降低百分比为35%,差异极为显著(p<0.05)。传统抗老药物中华麦饭石对受体随增龄下降具有对抗作用,但对KD值无影响。

【Abstract】 Neuronal nicotinic acetylcholine receptors (nAChRs) were involved in animal’s learning and memory activity as well as in human cognitive function of several degenerative diseases.In the past years, the research concerning the importance of neuronal nAChRs in cognitive function has attracted wide attention and there are some breakthroughs made in the central neuronal nAChRs due to the discovery of Kappa-Neurotoxins (K-NTs). However there are still much questions left concerning functions of nAChR in the central nervous system because of very limited K-NT available.In this paper, purified a-BGT(α-Bungarotoxin) and K-NT was used to prepared a model of crude membrane nAChRs from the heads of Drosophila melanogaster and a novel animal model based on the impaired learning and memory abilities These models were used to evaluate the anti-aging and intelligence-promoting functional food. Here are the main results:1. The preparation of crude membrance nAChRs from the heads of Drosophila melanogaster and its pharmacological properties.α -BGT and K-NT inhibit the binding of radiolabelled 1251-α-BGT to the crude membrane nAChR, the IC50 and Ki from the action of these

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