两种不同Duchenne型肌营养不良症模型鼠神经肌肉接头结构的比较  

Comparison of neuromuscular junction of two kinds of mouse models of Duchenne muscular dystrophy

作　　者：孔杰[1,2] 操基清[1] 杨娟[1] 陈菲[1] 李亚勤[1] 张成[1] 

机构地区：[1]中山大学第一附属医院神经科,广东省广州市510080 [2]广州中医药大学附属宝安区中医院脑病科,广东省深圳市518133

出　　处：《中国组织工程研究》2016年第5期657-663,共7页Chinese Journal of Tissue Engineering Research

基　　金：2015年卫生计生系统科研项目(201505021)~~

摘　　要：背景:Duchenne型肌营养不良症患者神经肌肉接头结构存在缺陷,主要表现为乙酰胆碱受体结构碎片化和突触后膜上棘状突起的减少,一直以来,公认这种结构上的缺陷是肌细胞结构破坏、肌纤维坏死所致。目的:探讨Duchenne型肌营养不良症模型鼠肌肉神经肌肉接头结构受损的原因。方法:引进雄性mdx小鼠和雄性dko小鼠(两者均为Duchenne型肌营养不良症模型),经基因鉴定后供实验使用,选用雄性C57BL/6小鼠为正常对照。采用苏木精-伊红染色检测模型鼠肌肉病理改变;应用免疫荧光染色显示神经肌肉接头结构。比较两种不同Duchenne型肌营养不良症模型鼠肌肉组织dystrophin表达、病理变化及神经肌肉接头结构差异。结果与结论:引进的模型鼠在基因型和蛋白表达层面均符合实验的要求,两种模型鼠神经肌肉接头上的乙酰胆碱受体数量有明显减少的趋势,尽管dko鼠肌肉较mdx鼠呈现出更为明显的炎性浸润和肌纤维破坏,但两者神经肌肉接头在结构上的受损并无明显差异,其乙酰胆碱受体的碎片化程度相似。这些证据表明,神经肌肉接头结构损伤和炎性病理反应是互相独立的事件,两者之间并无直接的关系。Dystrophin基因缺陷是导致乙酰胆碱受体结构碎片化的主要原因。BACKGROUND： Neuromuscular junction structure has defects in patients with Duchenne muscular dystrophy, mainly presenting acetylcholine receptor structure fragmentation and the reduction of spine-like processes on thepostsynaptic membrane. It is generally recognized that the structural defects are induced by structural damage of muscle cells and muscle fiber necrosis. OBJECTIVE： To explore the reasons of damage on neuromuscular junction in mouse models of Duchenne muscular dystrophy. METHODS： We introduced Duchenne muscular dystrophy models of male mdx mice and male Dko mice. After gene identification, they were used for tests. Male C57BL/6 mice were selected as normla controls. Hematoxylin-eosin staining was utilized to detect pathological changes in muscles. Neuromuscular junction structure was revealed using immunofluorescence staining. The differences in dystrophin expression, pathological features and neuromuscular junction structure were compared in mouse models of two kinds of Duchenne muscular dystrophy. RESULTS AND CONCLUSION： The introduced mouse models were accorded with the requirement of our experiment in aspects of genotype and protein expression levels. The number of acetylcholine receptor was apparently reduced in the neuromuscular junction of two kinds of mouse models. Although dko mouse muscles presented more obvious inflammatory infiltration and muscle fiber damage compared with mdx mice, but there was no significant difference in the damage to neuromuscular junction between them, and acetylcholine receptor fragmentation was identical. The evidence suggested that structural damage of neuromuscular junction and inflammatory pathological response are independent events. There is no direct relationship between them. Dystrophin gene deficiency is the main cause of the fragmentation of the acetylcholine receptor.

关 键 词：DUCHENNE型肌营养不良症 乙酰胆碱受体 组织工程 实验动物 神经损伤与修复动物模型 神经肌肉接头 

分 类 号：R318[医药卫生—生物医学工程]