原发性胆汁性肝硬化血清蛋白质谱的初步研究

目的筛选与原发性胆汁性肝硬化（PBC）相关的血清蛋白质分子标志物并建立指纹诊断模型。方法收集30例PBC患者血清,30例年龄、性别相匹配正常人,采用弱阳离子磁珠（MB-WCX）为检测介质,基质辅助激光解吸电离质谱（MADIL-TOF-MS）检测得到蛋白质谱,再应用基于磁珠的多维纳升级高效液相色谱及液体蛋白芯片指纹图谱（CLINPROT）系统比较两组血清蛋白质谱。结果在m/z100～10000范围内,检测出的129个蛋白峰中11个蛋白峰差异有统计学意义（P〈0.05）,其中m/z为4963.6、5805.33、1544.45、2292.16、2466.45、2845.85在PBC中表达上调,而m/z为3262.24、3191.2、4090.5、4072.43、3277.46的蛋白峰在PBC中下调;选择m/z为3262.24、3191.2这2个蛋白峰建立的蛋白芯片指纹图谱（CLINPROT）系统对PBC诊断的敏感性为90%,特异性为95%。结论筛选出的11个蛋白分子标志物可能与PBC有关,其中m/z4963在PBC中表达最明显;蛋白芯片指纹图谱（CLINPROT）系统可能对PBC的诊断具有一定临床意义。

Study on serum proteomics in primary biliary cirrhosis by using functionalized magnetic beads and MALDI-TOF-MS technique: a preliminary study

Objective To screen serum biomarkers in patients with primary biliary cirrhosis by using functionalized magnetic beads Weak Cation Exchange（WCX）and matrix-assisted laser desorption and ionization time-of-flightmass spectrometry（MALDI-TOF-MS）technique.Methods Proteomic spectra were generated by using mass spectroscopy in 60 cases,including 30 female cases of primary biliary cirrhosis that had been clinical confirmed with average aged（54.4±12.1）years,and 30 female cases of healthy control average aged（55.8±14.1）years.The mass spectrometry data obtained was analyzed using a new ClinProTools software.Results A total of 11 distinguished proteomic peaks were detected.Six peaks were up-regulated expression,including m/z 4 963.6、5 805.33、1 544.45、2 292.16、2 466.45、2 845.85,and five peaks were down-regulated expression,m/z 3 262.24、3 191.2、4 090.5、4 072.43、3 277.46.A diagnose proteomic pattern for primary biliary cirrhosis based on two selected proteomic peaks,3 262.24 and 3 191.2,could be built by using the ClinProTools software.Conclusion Functionalized magnetic beads MB-WCX and MALDI-TOF-MS offers a unique platform for the proteomic detection of primary biliary cirrhosis.A new ClinProTools software can detect the patients with primary biliary cirrhosis from normal controls.It also offers a noninvasive method to further study the proteomic changes in the development and progression of primary biliary cirrhosis.