结核性胸腔积液蛋白质组学初步研究

目的本实验利用二维凝胶电泳-肽质量指纹谱的蛋白质组学方法,对结核性、恶性和漏出性胸腔积液的相关蛋白质二维凝胶电泳（2-DE）图谱识别、鉴定,寻求结核性胸腔积液的特异蛋白质及其对临床诊断的意义。方法收集我院2010年5月～2011年6月收治的符合要求的胸腔积液患者50例,其中结核性胸腔积液20例,恶性胸腔积液17例,漏出性胸腔积液13例。利用二维凝胶电泳分离技术,基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析到相应的肽质量指纹谱（PMF）,最后借助Mascot软件结合NCBInr数据库进行检索鉴定相关结构蛋白质。结果通过对结核性、恶性和漏出性胸腔积液的蛋白质肽质量指纹图比较、分析发现3个存在明显差异蛋白,它们分别是：C1-抑制蛋白、甲状腺三级结构A链和补体C3b。C1-抑制蛋白在结核性胸腔积液高表达,在恶性胸腔积液中低表达。甲状腺素三级结构A链和补体C3b在恶性腔积液中高表达,在漏出液中低表达。结论利用二维凝胶电泳技术和基质辅助激光解吸电离飞行时间质谱技术对胸腔积液进行蛋白质组学研究,得到结核性、恶性和漏出性胸腔积液的蛋白质图谱,获得差异蛋白质,为结核性胸腔积液的鉴别诊断提供新的思路和方法。

Proteomic pilot study of tuberculosis pleural effusion.

Objective To seek useful protein biomarkers with diagnostic value by 2-DE and MALDI-TOF-MS technology in tuberculosis, malignant and transudatory-hydrothorax pleural effusion, and to identify and search tuberculosis pleural effusion specific protein for clinical diagnosis.Methods 50 examples of pleural effusion were collected in our hospital from May 2010 to June 2011, of which 20 examples were tuberculosis pleural effusion, 17 examples were malignant pleural effusion and other 13 examples were transudatory-hydrothorax.The proteins were isolated by two-dimensional gel electrophoresis （2-DE） and matrix-assisted laser desorption-time of flight-mass spectrometry （MALDI-TOF-MS） analysis were used to setup the corresponding peptide quality fingerprint spectra （PMF）, finally using the Mascot software combined NCBInr database retrieve to appraisal related structure protein. Results The PMF of tuberculosis, malignant pleural effusion and transudatory-hydrothorax were compared and analysised, and three different proteins were founded. They were Chain A, crystal structure of latent human C 1-inhibito; Chain A, tertiary structures of three amyloidogenic transthyretin variants;and implications for amyloid fibril formation; C 1-inhibiting protein had high expression in tuberculosis pleural effusion, and had low expression in malignant pleural effusion.Crystal structure of complement c3b and Chain a, tertiary structures of three amyloidogenic transthyretin had high expression in malignant pleural effusion, and low expression in transudatory-hydrothorax. Conclusion Using two-dimensional gel electrophoresis（2-DE） and matrix-assisted laser desorption-time of flight-mass spectrometry（MALDI-TOF-MS） to research pleura effusion protein, protein maps of tuberculosis, malignant pleural effusion and transudatory-hydrothorax and there different proteins are gathered. The results provide for us new train of thoughts and methods in tuberculosis pleural effusion differential diagnosis.