重组人乙酰胆碱受体α亚基1-210诱导实验性重症肌无力模型

发布时间：2018-05-04 02:17

本文选题：人乙酰胆碱受体a亚基1-210 + 实验性重症肌无力　；参考：《安徽医科大学》2006年硕士论文

【摘要】：背景：重症肌无力(MG)是一种主要累及神经肌肉接头突触后膜乙酰胆碱受体(AChR)，由乙酰胆碱受体抗体(AchR-ab)介导的自身免疫性疾病。发病率高达8～20／10万，并有逐年上升的趋势；且致残率高，严重危害人类健康，寻找有效治疗手段一直是研究者们努力的方向。实验性重症肌无力(EAMG)是其基础研究的动物模型；构建该模型的经典方法是由电鳗电器管中提取的纯化AchR作为免疫原，电鳗难以捕获，且提取过程复杂、费用昂贵，因此寻找构建该模型的新方法成为当务之急。AChR的主要生物学功能单位是α亚基，它具有高度的免疫原性，有研究证实刺激α亚基或α亚基的某些肽段，可以激活AChR特异反应性T细胞发生免疫反应。用含有免疫表位的α亚基的某些肽段作为免疫原诱导EAMG模型是近几十年来研究者们追求的目标；但可能是由于用作免疫原的肽段多在20个残基左右、免疫位点少、免疫原性较弱、须大剂量重复注射且肌无力存在时间短暂，不能成为理想的动物模型。AchRα亚基的N端细胞外区域1-210位残基，包含了ACh配体的结合位点和主要免疫原区，，是MG致病的关键区域，可以推测该肽段有可能成为有效免疫原。九十年代初Talib就应用基因技术分段克隆了hAChR α1-210片段，并在大肠杆菌中诱导表达；构建EAMG模型的知名专家Lennon则以该表达产物免疫Lewis鼠，结果成功诱导EAMG模型。随着分子生物学的发展，应用分子克隆和表达技术重组目的基因的方法已非常成熟；但国内外均未见使用类似方法诱导EAMG模型的报道。目的：我们首次参照Talib文献克隆并在大肠杆菌中诱导表达了hAChR α 1-210，然后用该表达产物免疫Lewis鼠诱导EAMG模型，希望为MG的基础研究提供一个简便、易行的实验动物模型。
[Abstract]:Background: myasthenia gravis (MG) is an autoimmune disease mediated by acetylcholine receptor (ache) receptor (ACHR), which mainly involves the postsynaptic membrane of neuromuscular junctions. The incidence rate is as high as 820 / 100, 000, and has a rising trend year by year, and the rate of disability is high, which seriously endangers human health. Finding effective treatment methods has been the direction of researchers' efforts. Experimental myasthenia gravis (EAMG) is an animal model for its basic research. The classical method to construct this model is to use purified AchR extracted from electric eel tube as immunogen, which is difficult to capture, and the extraction process is complex and expensive. Therefore, searching for a new method to construct this model has become an urgent task. The main biological functional unit of AChR is 伪 subunit, which has high immunogenicity. Some studies have confirmed that stimulating 伪 subunit or certain peptide segments of 伪 subunit. AChR specific reactive T cells can be activated to develop immune response. Using some peptides of 伪 subunit containing immune epitopes as immunogen to induce EAMG model has been pursued by researchers in recent decades, but this may be due to the fact that most of the peptides used as immune epitopes are about 20 residues and there are few immune sites. The immunogenicity is weak, the large dose of repeated injection and the short duration of myasthenia can not be an ideal animal model. The extracellular region of the N-terminal region of the AchR 伪 subunit is 1-210 residues, which contains the binding site and the main immunogen region of the ACh ligand. It can be inferred that the peptide may become an effective immunogen. In the early 1990s, Talib cloned the fragment of hAChR 伪 1-210 and induced expression in Escherichia coli, and Lennon, a well-known expert who constructed the EAMG model, immunized Lewis mice with the expression product, and the EAMG model was successfully induced. With the development of molecular biology, the methods of molecular cloning and expression to recombine the target gene have been very mature, but no similar methods have been reported at home and abroad to induce EAMG model. Objective: we cloned and induced the expression of hAChR 伪 1-210 in Escherichia coli with reference to Talib for the first time, and then immunized Lewis mice with this expression product to induce EAMG model, hoping to provide a simple and easy experimental animal model for the basic study of MG.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R746.1;R-332

【参考文献】