结核病患者和正常人血清蛋白质双向凝胶电泳差异比较  

Differential comparison of serum proteome between tuberculosis patients and healthy controls by using two-dimensional polyacrylamide gel electophoresis

作　　者：刘志辉[1] 刘玉美[1] 云径平[2] 罗强生[1] 尹小毛[1] 谭守勇[1] 刘健雄[1] 李昕洁[1] 

机构地区：[1]广东省广州市胸科医院,510095 [2]中山大学肿瘤防治中心,广州510060

出　　处：《广东医学》2008年第12期1991-1993,共3页Guangdong Medical Journal

基　　金：广州市科技计划资助项目(编号:2006J1-C0211)

摘　　要：目的初步探讨结核病患者与正常人之间血清蛋白质的差异,以期筛选出新的结核病实验诊断标志物。方法对10例菌阳肺结核(阳性组)、10例菌阴肺结核(阴性组)、10例肺外结核(肺外组)及10例正常人(正常组)分组混合血清应用17cmpH4～7IPG胶条进行双向凝胶电泳(two-dimensional electrophoresis,2-DE),每组平行电泳3块胶条。电泳结束后对2-DE胶条进行考马斯亮蓝染色,染色后用Umax powerlook2000扫描仪以投射方式、300dpi分辨率扫描获取2-DE图谱,然后用2-DE分析软件Image Master2D Platium5.0进行分析。结果在正常组、阳性组、阴性组、肺外组中,分别平均可以检测到310±64、307±33、296±54、267±23个清晰可见的蛋白质点。正常组与阳性组、阴性组、肺外组的匹配率分别为(74±2)%、(76±2)%和(68±5)%。通过对4组2-DE图谱比较,阳性组与正常组间有21个差异蛋白质点(其中9个上调),阴性组与正常组间有18个差异蛋白质点(其中8个上调),肺外组与正常组间有24个差异蛋白质点(其中7个上调)。将阳性组、阴性组、肺外组合并为结核组与正常组比较有3个差异蛋白质点(其中2个上调)。结论结核病患者和正常人血清蛋白质存在差异,有望从血清中筛选出新的结核病实验诊断标志物。本研究建立了两者间血清蛋白质组差异蛋白质的初步模型,但有待进一步验证并进行差异蛋白质鉴定。Objective To investigate the difference of serum proteome between tuberculosis patients and healthy controls and to screen out novel laboratory diagnosis marker of tuberculosis. Methods Samples of peripheral blood were collected from patients with bacterial positive pulmonary tuberculosis（ ＋ group）, patients with bacterial negative pulmonary tuberculosis（ - group ）, patients with ex - pulmonary tuberculosis （ ex - pulmonary group ）, healthy controls （ control group） ,with 10 patients in each group. Serum samples of each group were mixed respectively by different proportion according to the protein concentration of the samples. Two - dimensional polyacrylamide get electrophoresis （ 2 - DE） was performed on immobilized pH gradient rod gels（pH4 -7, 17 cm） and each mixed sample was tested three times simuhaneously. The gels were stained using colloidal Coomassie Blue, and scanned using Umax powerlook2000 scanner to acquire images of 2 - DE gel in the condition of projection mode and 300dpi resolution. Analysis of the image was performed with ImageMaster 2D Platium.5.0. Results 310 ＋ 64,307 ＋ 33,296 ±54 and 267 ± 23 of clear protein spots were detected respectively in the 2 -DE gel of control group, ＋ group, -group and ex -pulmonary group. The matching rates with control group matched ＋ group and- group and ex- pulmonary group, were 74% ± 2% ,76%± 2% and 68% ± 5% respectively. Making a comparison among four groups, there were 21, 18, and 24 differentially expressed proteins respectively between control group and ＋ group, - group, ex - pulmonary group. And there were 9, 8, 7 up - regulation expression proteins in ＋ group, - group and ex - pulmonary group. There were 3 differentially expressed proteins including 2 up - regulation expression proteins in tuberculosis group based on ＋ group and - group and ex - pulmonary group. Conclusion There are differentially expressed proteins in serum between tuberculosis patients and healthy controls and that novel laboratory diagnosis m

关 键 词：结核 双向凝胶电泳 蛋白质组 血清蛋白质组学 

分 类 号：R684.3[医药卫生—骨科学] R735.7[医药卫生—外科学]