应用蛋白质组学技术对脂肪肝患者血清标志物的筛选

Screening serum biomarkers of fatty liver disease using proteomic technology

目的:用纳米磁珠与基质辅助激光解析离子化飞行时间质谱(MALDI-TOF-MS)技术检测脂肪肝患者血清蛋白质组,筛选候选标志物并建立诊断模型,初步探讨所建立的诊断模型在脂肪肝早期诊断中的临床意义.方法:95例中度和76例轻度脂肪肝患者均经B超证实,99名健康人血清.用WCX(弱阳离子交换型)纳米磁珠检测各血清标本获得血清蛋白指纹图谱.用Biomarker Wizard软件分析所获得的蛋白图谱找出差异蛋白,再用Biomarker Patterns建立诊断模型.扩大样本量,通过盲法分析进一步验证诊断模型的可靠性.结果:初始建模时,建立了由m/z为7626.2、24147.7、6118.2这3个差异蛋白峰组成的中度脂肪肝诊断模型,其敏感性为90.2%,特异性为93.9%.扩大样本盲法验证的敏感性为90.9%,特异性为92.0%.另选取了m/z为7626.2、3286.0、2760.7、2543.1、2746.1这5个差异蛋白峰组成的模型,能把轻度从中度脂肪肝患者中区分出来;选取m/z为7626.2、11721.0、24141.7、8529.1、9042.7这5个差异蛋白峰组成的模型,能将轻度脂肪肝患者从健康对照中区分出来.结论:蛋白质组学技术可筛选出有意义的血清生物标志差异蛋白.由三个差异表达蛋白及其特定组合构成的诊断模型可以区分中度脂肪肝患者与健康人,由五个特定组合构成的差异表达蛋白的模型能区分轻度、中度脂肪肝患者及区分轻度脂肪肝患者与健康对照.

AIM： To analyze the serum proteomic pattern of the fatty liver disease （FLD） patients, to build a diagnostic model and to evaluate its clinic significance using magnetic bead and MALDI-TOF-MS. METHODS： WCX magnetic bead and MALDI- TOF-MS were used to detect the serum proteomic patterns of 95 modest FLD patients, 76 mild FLD patients and 99 healthy persons （HP）. Biomarker Wizard software was used to analyze protein peaks for significant difference. The model was then built by Biomarker Patterns Software and was further evaluated in blind test for reliability. RESULTS： A diagnostic pattern for modest FLD from HP using m/z 7626.2, 24147.7, 6118.2 was established with 90.2% sensitivity and 93.9% specificity. A sensitivity of 90.9% and a specificity of 92.0% in blind test were obtained. One diagnostic pattern using m/z 7626.2, 3286.0, 2760.7, 2543.1, 2746.1 was established to differentiate mild from modest FLD; the other diagnostic pattern using m/z 7626.2, 11721.0, 24141.7, 8529.1, 9042.7 was established to differentiate mild FLD from HP. CONCLUSION： Proteomic technology can significantly identify novel significant biornarkers in the serum. The diagnostic pattern consisting of three differential protein peaks can discrimi- nate the modest FLD from HP. The diagnostic pattern consisting of five specific differential protein peaks can differentiate mild from modest FLD as well as mild FLD from HP.