慢性阻塞性肺疾病患者外周骨骼肌萎缩蛋白质组初步分析

Preliminary proteomic analysis of peripheral skeletal muscle atrophy in chronic obstructive pulmonary disease

目的初步分析慢性阻塞性肺疾病（COPD）患者外周骨骼肌萎缩蛋白质组。方法研究对象为2010年2_7月昆明医学院第一附属医院收治的16例COPD患者（COPD组）和同期收治的年龄匹配的8例骨折患者（对照组）。根据患者体质指数（BMI）、去脂肪指数、股四头肌的周径、股四头肌最大主动收缩力结果，COPD组又分为：（1）骨骼肌萎缩组8例：男6例，女2例；平均（70±8）岁；（2）骨骼肌非萎缩组8例：男5例，女3例；平均（74±8）岁。对照组男6例，女2例；平均（72±6）岁。利用二维凝胶电泳分离出各组受试者股四头肌外侧肌组织的总蛋白；通过PDQuest图像软件比较电泳图谱中的异常蛋白点，识别差异表达蛋白；应用质谱仪得到相应的肽质量指纹谱；然后搜索数据库鉴定部分差异蛋白点。结果获得分辨率和重复性均较好的双向凝胶电泳图谱，并鉴定出可能与COPD肌萎缩发生密切相关的12种蛋白，其中8种结构蛋白（心肌α-肌动蛋白解离亚型c、心肌肌球蛋白调节轻链-2、肌红蛋白及其亚型、心肌β-肌球蛋白解离亚型7c多肽、骨骼肌α-肌动蛋白、心肌仪．肌动蛋白解离亚型b、血红蛋白珠蛋白1α链、肌球蛋白轻链-6B），4种功能蛋白（烯醇化酶1-A链、慢骨骼肌肌钙蛋白T1解离亚型a、碳酸酐酶、慢骨骼肌肌钙蛋白T解离亚型b）。结论COPD患者常并发有明显外周骨骼肌萎缩，原因可能与外周骨骼肌结构蛋白和功能蛋白数量及质量的改变有关。

Objective To conduct a preliminary proteomic study of chronic obstructive pulmonary disease （COPD） with peripheral skeletal muscle atrophy. Methods A total of 16 COPD patients and 8 aged-matched persons because of bone fractures were recruited in First Hospital Affiliated to Kunming Medical College from February to July in 2010. According to body mass index, fat free mass index, quadriceps femoris perimeter and quadriceps femoris active contraction, they were divided into those with muscle atrophy （ group A, n = 8 ） and those without （ group B, n = 8 ）. There were 6 males and 2 females with an average age of （70 ± 8 ） years in the group A and 5 males and 3 females with an average age of （74 ±8） years in the group B. And the control group had 6 males and 2 females with an average age of （72 ±6） years. All samples of total quadriceps protein were separated by two-dimensional gel eleetrophoresis. The abnormal protein points on electrophoresis were compared by PDQuest image software. And the differential protein expression was detected. Then the corresponding peptide quality fingerprint spectrum was analyzed by mass spectrometer. Finally the differential protein points were partially detected by a search of database. Results The two-dimensional gel electrophoresis yielded an excellent profile of resolution and repeatability. And 12 proteins likely to cause skeletal muscle atrophy in COPD were identified. Among them, 8 proteins belonged to structural proteins （actin alpha cardiac muscle isoform CRAc, myosin regulatory light chain 2, ventricular/cardiac muscle isoform, myoglobin isoform myosin heavy polypeptide 7 cardiac muscle beta isoform CRA_c, actin, alpha skeletal muscle, actin alpha cardiac muscle isoform CRA_b, hemoglobin alpha 1 globin chain ＆ myosin light chain 6B） and 4 proteins were of functional proteins （chain A, crystal structure of human enolase ; troponin T, slow skeletal muscle isoform alpha, carbonate anhydrase, troponin T ＆ slow skeletal muscle isoform b）. Conclusions COPD patients are often accompanied with obvious peripheral skeletal muscle atrophy. It may be caused by quantitative or qualitative changes of peripheral skeletal structural and functional proteins.