肌营养不良症模型鼠神经元型一氧化氮合酶抑制蛋白的表达及定位  

Expression and localization of protein inhibitor of neuronal nitric oxide synthase in mice with muscular dystrophy

作　　者：毕胜[1] 王德生[1] 韩丽丽[1] 王雪婷[1] KAO Yanlin 

机构地区：[1]哈尔滨医科大学第一临床医学院神经内科 [2]澳大利亚新南威尔士大学悉尼儿童医院Westfield研究室

出　　处：《中国临床康复》2004年第22期4450-4453,共4页Chinese Journal of Clinical Rehabilitation

基　　金：博士后基金(LRB00071)~~

摘　　要：目的:观察肌营养不良症模型鼠(mdx)肌肉组织神经元型一氧化氮合酶抑制蛋白(proteininhibitorofnNOS,PIN)的表达及定位,以及与dys-trophin复合物及神经元型一氧化氮合酶(nNOS)的关系。方法:取mdx鼠(18只,实验组)和C57BL/10ScSn鼠(15只对照组)骨骼肌作常规染色,比较其组织学改变;同时对mdx鼠,C57BL鼠骨骼肌作Northernblot和固定化蛋白质印迹法(Westernblot)及免疫组化分析。结果:Northernblot分析发现,mdx鼠肌肉组织PINmRNA显著高于C57BL/10ScSn鼠,对照组骨骼肌有少量PINmRNA表达,其杂交斑点A值为1352.4±219.7,mdx组杂交斑点灰度A值为2724.2±394.6,两者差异有非常显著性意义(t=11.985,P<0.05);固定化蛋白质印迹法(Westernblot)显示,PIN蛋白的杂交信号在mdx组与对照组均出现在约8ku处,两者的吸光度平均值(A)几乎相等,mdx组的杂交斑点灰度A值为2378.2±486.7,对照组PIN蛋白的杂交斑点灰度A值为2438.5±494.9,两者差异无显著性意义(t=0.352,P>0.05);免疫组化及激光共聚焦扫描显微镜均揭示,在C57BL/10ScSn鼠肌纤维中,PIN蛋白在肌膜、周边核附近呈强阳性表达,肌质中弱表达,而mdx鼠的PIN蛋白在肌膜及中心核附近呈强阳性表达,细胞质中亦弱表达;PIN于mdx鼠肌纤维表达明显,而nNOS及dystrophin复合物表达明显减少?AIM:To observe the expression and localization of protein inhibitor of neurona l nitric oxide synthase(nNOS)(PIN) in the muscle tissues of rats with muscular d ystrophy(mdx),and to explore the relation of PIN with the dystrophin complex and nNOS. METHODS:Routine staining was performed in the skeletal muscles of 18 mdx mice( experimental group) and 15 C57BL/10ScSn mice(control group),and the histological changes were compared between the two groups;meanwhile Northern blot,Western bl ot and immunohistochemical analysis were also performed in the two groups. RESULTS:Northern blot showed that the PIN mRNA in muscle tissue of the mdx rat s was significantly higher than that of C57BL/10ScSn rats,there were a few PIN m RNA expressions in the skeletal muscles in the control group.The hybrid spot abs orbance(A) values were significantly different between the control group(1 352.4 ±219.7) and mdx group(2 724.2±394.6)(t=11.985,P< 0.05).Western blot showed tha t the hybrid signals of PIN protein were observed at 8 ku in both groups,and the mean A values were almost the same in the two groups.The hybrid spot A values w ere also not significantly different between the mdx group(2 378.2±486.7) and t he control group (2 438.5±494.9)(t =0.352,P >0.05).Immunohistochemical analy sis and the confocal laser scanning microscope both showed that in the muscle fi bers of C57BL/10ScSn rats, the strong positive expression of PIN protein was loc alized at the sarcolemma and peripheral nuclei,while weak expression was found i n muscular substance.By comparison,the strong positive expression of PIN protein was more concentrated around the sarcolemma and central nuclei,while weak expre ssion was found in cytoplasm in mdx mice.The presence of PIN protein expression in muscles of mdx mice was evident,while the expressions of nNOS and dystrophin complex were obviously reduced. CONCLUSION:PIN is not an integral component of the dystrophin complex inside s keletal muscle fibers. PIN expression in muscles from mdx mice is controll

关 键 词：肌营养不良 一氧化氮合酶 抑制蛋白 印迹法 蛋白质 

分 类 号：R746.2[医药卫生—神经病学与精神病学]