小分子差异蛋白在HBV相关性肝病患者血清中的SELDI-TOF-MS筛选  

Screening of differentially expressed low-abundance proteins among serum samples from patients with different HBV-related hepatic diseases by SELDI-TOF-MS

作　　者：宁秋悦[1] 吴继周[1] 李国坚[1] 臧宁[1] 胡蝶飞[1] 吴健林[1] 陈茂伟[1] 万裴琦[1] 

机构地区：[1]广西医科大学第一附属医院感染性疾病科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2011年第2期143-150,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30760219~~

摘　　要：目的:探讨乙型肝炎病毒(HBV)相关性肝病患者血清中小分子差异蛋白在HBV感染后病情发展过程中的意义.方法:采用蛋白芯片及表面增强激光解析电离飞行时间质谱(surface-enhanced laserdesorption/ionization time-of-flight massspectrometry,SELDI-TOF-MS)技术对已用乙腈去除高丰度蛋白的正常对照(NC)、乙型肝炎病毒携带者(ASCs)、慢性乙型肝炎(CHB)、肝硬化(LC)及原发性肝细胞癌(HCC)患者术前血清进行检测,筛选各自的血清小分子差异表达蛋白,并分别建立诊断模型.在蛋白数据库expasy寻找相关差异蛋白信息,对差异蛋白峰的可能结构及功能进行评价.结果:与NC组比,ASCs组有63个蛋白质波峰的强度值存在统计学差异(P<0.05),其中29个上调,34个下调;CHB组有57个,其中21个上调,36个下调;LC组有68个,其中33个上调,35个下调;HCC组有74个,其中28个上调,46个下调;通过对比分析,发现在4个病例组表达均为上调的m/z为15889.8,蛋白峰强度值在NC<ASC<CHB<LC组,HCC组较CHB和LC组低;11742.2蛋白峰强度值在NC<ASC<CHB<LC组和NC<ASC<CHB<HCC组,在LC和HCC组最高,用此蛋白峰诊断HBV感染相关性LC的灵敏度和特异度分别为90%和86.67%,诊断HCC的灵敏度和特异度分别为93.33%和83.33%.结论:成功去除高丰度蛋白和应用SELDI-T O F-M S技术筛选H B V感染相关性肝病患者血清中小分子差异蛋白,m/z为8709.7、13759.8、14004.0、15361.89、16072.3、2746.8、3449.1、3941.06、4098.3、9445.5的10个蛋白峰可能与HBV感染有关;m/z为15889.8的蛋白峰可能成为H B V感染后进展为LC早期诊断的标志物,而m/z为11742.2的蛋白峰也许是HBV相关性LC或HCC的一个重要标志.AIM： To screen differentially expressed low- abundance proteins among serum samples from patients with different HBV-related hepatic diseases and to evaluate their possible value in the diagnosis of these diseases. METHODS： The surface-enhanced laser de- sorption or ionization time-of-flight mass spec- troscopy （SELDI-TOF-MS） was used to screen differentially expressed proteins among serum samples, in which high-abundance proteins had been removed with acetonitrile, collected from patients with asymptomatic chronic hepatitis B （ASC）, chronic hepatitis B （CHB）, liver cirrho- sis （LC）, hepatocellular carcinoma （HCC）, and normal controls. Diagnostic models for each disease were then established with differentially expressed proteins. Protein databases were searched to predict the possible structure and function of differentially expressed proteins. RESULTS： Compared with the normal control group, 63 differentially expressed protein peaks were detected in the ASC group, of which 29 were up-regulated and 34 down-regulated （P 〈 0.05）; 57 in the CHB group, of which 29 up- regulated and 34 down-regulated; 68 in the LC group, of which 33 up-regulated and 35 down- regulated; and 74 in the HCC group, of which 28 up-regulated and 46 down-regulated. A peak with a m/z of 15 889.8 corresponded to a protein whose expression was up-regulated gradually in an order of healthy controls, ASC, CHB and LC patients, and its expression level in the HCC group was lower than those in the CHB group and LC group. The expression of a protein with a m/z of 11 742.2 was higher in the LC group and HCC group than in other groups, and its expression was gradually increased in an order of healthy controls, CHB, and LC patients, or in an- other order of healthy controls, CHB, and HCC patients. The sensitivity and specificity using the protein peak at 11 742.2 for diagnosis of LC were 90% and 86.67% and for HCC were 93.33% and 83.33%, respectively. CONCLUSION： Ten protein peaks with m/z values of 8 709.7, 13 759.8, 1

关 键 词：乙型肝炎病毒携带者 慢性乙型病毒性肝炎 肝炎肝硬化 原发性肝细胞癌 低丰度蛋白 表面加强激光解吸电离一飞行时间质谱 

分 类 号：R512.62[医药卫生—内科学]