抗肌肉特异性激酶单克隆抗体对大鼠重症肌无力的诱导

发布时间：2018-02-03 11:21

  本文关键词： 肌肉特异性激酶 重症肌无力 单克隆抗体 免疫荧光 透射电镜　出处：《延边大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过建立Lewis大鼠实验性自身免疫性重症肌无力(myasthenia gravis,MG)模型(experimental autoimmune MG, EAMG),以此研究肌肉特异性激酶(muscle-specific kinase, MuSK)抗体(muscle-specific kinase antibody, MuSKAb)和MG发病机制的关系。 方法：选取10只雌性Lewis大鼠,随机分为实验组和对照组。采用被动免疫的方法,将MuSK单克隆抗体通过腹腔注射的方式给予实验组大鼠,对照组大鼠采用同样的方法给药,每2小时观察1次大鼠的饮食饮水情况以及有无临床症状出现,并随时记录大鼠的体重变化。给药96小时后,迅速处死两组大鼠,并取下大鼠的肋间肌、腓肠肌和胫前肌,与-80℃冰箱中保存。之后对于取下的肌肉组织进行免疫荧光实验,用以观察抗MuSK单克隆抗体和MuSK蛋白结合情况和C3补体激活情况。同时做透射电镜实验,用以观察神经肌肉接头处(NMJ)的形态学变化。 结果：在给药96小时后实验组与对照组相比,未出现抓握和嘶叫无力的明显临床症状。经过免疫荧光实验后发现,实验组肋间肌、腓肠肌和胫前肌的细胞膜表面出现明显的黄绿色荧光和红色荧光物质,黄绿色荧光说明MuSK单克隆抗体和跨膜蛋白MuSK发生特异性结合,红色荧光说明C3补体抗体和C3发生特异性结合。而对照组无荧光出现。透射电镜观察显示实验组大鼠运动终板突触后区域简单化,突触后膜皱褶退化,分级减少并短缩,突触间隙略增宽。而实验组无明显变化。 结论：将MuSK单克隆抗体注射给大鼠后未能引起肌无力的临床症状,但在细胞膜处发现,MuSKAb和MuSK发生特异性结合,并且能够激活C3补体,同时NMJ发生了超微结构形态学变化。抗MuSKAb的致病性仍需进一步研究。
[Abstract]:Objective: to establish myasthenia gravis of experimental autoimmune myasthenia gravis in Lewis rats. Experimental autoimmune (EAMG). To study muscle-specific kinase. The relationship between muscle-specific kinase antibody (MuSKAb) and the pathogenesis of MG. Methods: ten female Lewis rats were randomly divided into experimental group and control group. The MuSK monoclonal antibody was injected intraperitoneally into the experimental group by passive immunization. Rats in the control group were treated with the same method. The diet and drinking water status and clinical symptoms were observed once every 2 hours, and the weight changes of the rats were recorded at any time. 96 hours after administration of the drug. The intercostal muscle, gastrocnemius muscle and tibial anterior muscle of the rats were quickly killed and stored in refrigerator at -80 鈩，

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