利用SELDI-TOF-MS技术研究巨细胞病毒感染致婴儿肝炎综合征的血清蛋白标志物

Identif ication of serum biomarkers in human cytomegalovirus infected hepatitis infants by SELDI-TOF-MS

目的:用表面增强激光解吸离子化飞行时间质谱(SELDI-TOF-MS)技术研究人巨细胞病毒(hCMV)感染致婴儿肝炎综合征患儿血清蛋白质谱的差异表达,寻找特异性蛋白标志物,为研究hCMV感染引起婴儿肝炎综合征的蛋白质组学发病机制奠定基础.方法:实验组为hCMV感染所致的婴儿肝炎综合征患儿20例;对照组Ⅰ为感染hCMV但未发生婴儿肝炎者5例;对照组Ⅱ为hCMV感染阴性的婴儿肝炎10例;对照组Ⅲ为健康对照10例.采集全血,分离血清,用SELDI-TOF-MS技术和WCX2芯片检测各组蛋白质谱的表达.结果:hCMV感染引起的婴儿肝炎综合征组与其他对照组比较,血清中有4个蛋白质表达水平发生明显变化,分别为Mr4349.8、Mr5808.7、Mr7935.6和Mr8885.9的蛋白峰.hCMV感染的两个组与未感染的对照组婴儿比较,血清中共有5个蛋白质表达水平发生明显变化,分别为Mr3266.8、Mr5638.5、Mr5909.1、Mr7771.4和Mr15835.6的蛋白峰.hCMV感染但未发生婴儿肝炎组与其他组婴儿比较,血清中有2个蛋白质表达水平发生明显变化,分别为Mr4600.1和Mr5704.3的蛋白峰.两个婴儿肝炎组与肝功能正常的另两组婴儿比较,血清中有4个蛋白质表达水平发生明显变化,分别为Mr7567.0、Mr13744.8、Mr15100.7和Mr15915.0的蛋白峰.结论:一些特定的血清蛋白质分子如肝再生增强因子、前白蛋白和结合珠蛋白是与肝功能密切相关的因子;细胞因子β-防御素31,β-防御素8和巨细胞源性趋化因子等,可能与hCMV感染、感染后发病及感染后保护相关.

AIM： To identify the serum protein biomarkers in infants with hepatitis syndrome resulted from congenital human cytomegalovirus （hCMV） infected using surface enhanced laser desorption/ionization-time of flight mass spectrometry （SELDI-TOF-MS）. METHODS： Serum samples were collected from 20 hCMV infected hepatitis infants and 25 controls. Of the 25 infants, 5 were infected with hCMV but without hepatitis, 10 with hepatitis but no hCMV infection, and 10 were healthy infants. The proteomic expression in the serum was detected by WCX2 chips and SELDI-TOF-MS. RESULTS： Fifteen protein peaks were distinctly different among the four groups in the mass range of Mr 2 000-20 000. Of the 15 peaks, four at/14r4 349.8,/14r 5 808.7, Mr 7 935.6 and Mr 8 885.9 were significantly different between the congenital hCMV infected hepatitis infants and the controls. Five peaks were distinctly upregulated in the infants with hCMV infection （M,3 266.8, Mr 5 638.5, Mr 5 909.1, Mr 7 771.4 and Mr 15 835.6） as compared with those in the infants without hCMV infection. In comparison with other infants, two proteins at Mr 4 600.1 and Mr 5 704.3, respectively, were upregulated in the infants with hMCV infection but no hepatitis. Four protein peaks were markedly different （Mr 7 567.0, Mr 13 744.8, Mr 15 100.7 and Mr15 915.0） between the infants with hepatitis and other controls. CONCLUSION： Some specific serum protein such as augmenter of liver regeneration, prealbumin, and haptoglobin were closely related to liver function. Cytokines such as beta-defensin 31, 8, and macrophage-derived chemokine, and etc were involved in the hMCV infection and the pathogenesis hMCV-induced hepatitis in infants.