肌肉中糖原含量与形态测定和糖原脱支酶活性检测诊断ⅢA型糖原累积症  

Diagnosis of glycogen storage disease type Ⅲ A by detecting glycogen debranching enzyme activity, glycogen content and structure in muscle

作　　者：王薇[1] 魏珉[1] 宋红梅[1] 邱正庆[1] 张为民[2] 吴晓燕[1] 卢超霞[3] 祁俊明[1] 景红[1] 李凡[1] 

机构地区：[1]中国医学科学院 北京协和医学院 北京协和医院儿科,100730 [2]中国医学科学院 北京协和医学院 北京协和医院中心实验室,100730 [3]中国医学科学院 北京协和医学院 北京协和医院基础医学院遗传室,100730

出　　处：《中华儿科杂志》2009年第8期608-612,共5页Chinese Journal of Pediatrics

基　　金：国家重点基础研究发展计划（973计划）（2005CB522507）;“十一五”国家科技支撑计划项目（2006BA105A07）;北京市科技计划项目（D0906005040491）;北京协和医院临床重点项目基金 志谢 The International Collaborative Genetics Research Training Program（ICGRTP,NIH D43 TW06176）项目为我们提供人员培训机会.Duke University Medical Center Glycogen Storage Disease Laboratory慷慨提供酶学检测方法.北京协和医院神经科、外科、手术室的支持与帮助

摘　　要：目的在我国建立糖原累积症（GSD）ⅢA的酶学诊断方法，并测定中国人群的正常值范围。方法肌肉标本来自正常对照组35例、GSD ⅢA组12例和其他肌病组9例。GSD ⅢA患者均发现明确的AGL基因致病突变。制备肌肉匀浆后，以淀粉葡萄糖苷酶以及磷酸化酶分别降解匀浆中的糖原，通过测定葡萄糖产量以明确糖原含量以及糖原形态；以极限糊精为底物，测定匀浆中的糖原脱支酶（GDE）活性。结果（1）GSD ⅢA患者组的肌肉中糖原含量显著高于另外两组，葡萄糖-1-磷酸与总葡萄糖的比率（G1P／G）以及GDE活性均显著低于另外两组（P〈0．01），其他肌病组与正常对照组三项指标的差异均无统计学意义。（2）正常值范围：肌肉中糖原含量0．31％～0．43％，G1P／G为22．37％～26．43％，GDE活性0．234～0．284μmol／（g·min）。（3）三项指标综合诊断GSD ⅢA的酶学诊断方法与基因诊断方法诊断效果相同，并且两种方法的敏感性均为91．7％，特异性均为100．0％。结论首次建立了GSD ⅢA的酶学诊断及糖原含量、糖原形态测定方法，确定r中国人群的正常值范围，可于临床推广应用。Objective Glycogen storage disease type Ⅲ （GSD Ⅲ ） is an autosomal recessive disease caused by glycogen debranching enzyme （GDE） gene （AGL gene ） mutation resulting in hepatomegaly, hypoglycemia, short stature and hyperlipidemia. GSD Ⅲ A, involves both liver and muscle, and accounts for up to 80% of GSD Ⅲ. The definitive diagnosis depends on either mutation analysis or liver and muscle glycogen debranching enzyme activity tests. This study aimed to establish enzymologic diagnostic method for GSD m A firstly in China by detecting muscular GDE activity, glycogen content and structure and to determine the normal range of muscular GDE activity, glycogen content and structure in Chinese children. Method Muscle samples were collected from normal controls （male 15, female 20; 12 -78 years old）, molecularly confirmed GSD mA patients （male 8, female 4, 2 -27 years old） and other myopathy patients （male 9, 2 - 19 years old ）. Glycogen in the muscle homogenate was degraded into glucose by amyloglucosidase and phosphorylase respectively. The glycogen content and structure were identified by glucose yield determination. The debranching enzyme activity was determined using limit dextrin as substrate. Independent samples Kruskal-Wallis H test, Nemenyi-Wilcoxson-Wilcox test, and Chi-square test were used for statistical analyses by SPSS 11.5. Result （ 1 ） GSD Ⅲ A patients＇ glycogen content were higher, but G1P/G ratio and GDE activity were lower than those of the other two groups （P 〈0. 01 ）. In all of the three parameters, there were no significant difference between normal controls and other myopathy patients. （2） The range of normal values： glycogen content 0. 31% -0. 43% , G1P/G ratio 22. 37% -26.43%, GDE activity 0. 234 - 0. 284 μmol/（ g. min）. （ 3 ） Enzymologic diagnostic method had a power similar to that of gene analysis in diagnosis of GSD-Ⅲ A patients. The sensitivity and specificity of enzymologic diagnostic method and mutation detection were 91.7% and 100

关 键 词：糖原贮积病Ⅲ型 诊断技术和方法 糖原脱支酶系统 

分 类 号：R686[医药卫生—骨科学]