壮族腰椎间盘突出症血瘀证血清蛋白质指纹图谱诊断模型初步研究  

Preliminary research of serum protein fingerprinting model for the diagnosis of the blood stasis of lumbar intervertebral disc protrusion in Zhuang population

作　　者：许建文[1] 钟远鸣[1] 尹利军[1] 胡兵[2] 吕建珍[2] 张家立[1] 李智斐[1] 陈静[1] 李桂焕[1] 

机构地区：[1]广西中医药大学第一附属医院骨科,南宁530023 [2]广西中医药大学,2009级研究生南宁530001

出　　处：《中国矫形外科杂志》2013年第19期1985-1990,共6页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81060300);广西自然科学基金项目(桂科自2011GXNSFA018181);广西高等学校优秀人才资助项目{桂教财基[2010]92号}

摘　　要：[目的]应用表面增强激光解吸/电离飞行时间质谱技术(surface-enhanced laser desorption/inionation time of flight mass spectrometry,SELDI-TOF-MS)筛选腰椎间盘突出症(lumbar intervertebral disk protrusion,LIDP)壮族血瘀证患者血清差异表达蛋白,建立壮族LIDP血瘀证的血清蛋白指纹图谱模型,并验证血清中的差异蛋白峰。[方法]按既定统一标准选取30例壮族LIDP血瘀证患者作为观察组,然后将同期就诊并与观察组患者年龄、性别等基线资料相近的30例汉族LIDP血瘀证患者作为对照组。均按同样方法提取60例患者外周血的血清样本,采用SELDI-TOF-MS及蛋白质芯片技术检测并绘制蛋白质质谱图并用Biomarker Wizard软件识别蛋白峰信息,再运用SPSS16.0软件及Biomarker Patterns软件对数据进行统计分析处理,建立壮族LIDP血瘀证的血清诊断模型,然后对模型进行双盲验证并通过ExPASy数据库对相关差异蛋白进行蛋白检索。[结果]在两组组间有75个明显表达差异的蛋白质峰(P<0.05),用其中三个蛋白即可对LIDP壮族血瘀证和汉族血瘀证进行正确区分,其准确性分别为LIDP壮族血瘀证100%、汉族血瘀证100%;另外选取单独的68例样本对模型进行双盲验证测试,结果准确性分别为LIDP壮族血瘀证94%、LIDP汉族血瘀证83%;对建立决策树模型的3个蛋白通过分子量检索推测,显示这3个蛋白中有1个(4 011.45)在LIDP壮族血瘀证组高表达、有2个(1 191.53、1 412.52)在LIDP汉族血瘀证组高表达。[结论]壮族LIDP血瘀证患者血清中存在异常表达的蛋白质;由3个差异蛋白组成的血清蛋白质指纹图谱模型可作为壮族LIDP血瘀证的血清标志物诊断模型。[ Objective] Surface - enhanced laser desorption/inionation time of flight mass spectrometry （ SELDI - TOF - MS） technology was undertaken to screen the diverse expression proteins of the peculiar serum of blood stasis syndrome of LIDP in Zhuang, establish the serum protein model of fingerprint atlas of the blood stasis syndrome of LIDP in Zhang, and confirm the peak value of correlated serum proteins. [ Methods]Thirty cases were chosen by identified unify standard as the observation group of the blood stasis of LIDP in Zhuang. Thirty cases were chosen by the same time of treatment and the same ages , sex, baseline date to the observation group as the control group of the blood stasis of LIDP in Han. Serum samples of peripheral blood from the 60 patients were collected through the same method. SELDI - TOF - MS and ProteinChip technology were employed to detect and draw the protein~ mass spectrometry. Then the protein peak values were identified by Biomarker Wizard software. The data were analyzed statistically by SPSS 16. 0 and dealt by Biomarker Patterns. At last, serum diagnosis model of the blood stasis syndrome of LIDP in Zhuang was established and the model was tested and verified through double blind test method. The related different proteins were searched by ExPASy data. [ Results ] Seventy - five differentially expressed protein peaks were found be- tween the two groups （ P 〈 0. 05 ） . Three of the proteins were enough for distinguish the blood stasis of LIDP in Zhuang and thatin Han, the accuracy were all 100% in blood stasis of LIDP in Zhuang and Han. The model was tested and verified through double blind test method by selecting another 68 samples , the ac- curacy was 93. 939 % of the blood stasis of LIDP in Zhuang and 83% in Han. We set up the model of decision tree inference of 3 proteins by molecular weight retrieval. It showed that 1 （4 011.45） was highly expressed of the blood stasis of LIDP in Zhuang and other 2 （ 1 191.53, 1 412. 52） were relatively highlyexpressed in Han. [ C

关 键 词：腰椎间盘突出症 证型 生物学 蛋白质组学 蛋白质芯片SELDI—TOF—MS 

分 类 号：R274[医药卫生—中医骨伤科学]