肾移植排斥反应患者血清蛋白质谱的差异性研究

目的 研究肾移植排斥反应患者的血清蛋白质谱的差异性,检测肾移植排斥反应早期诊断的特殊血清多肽标记物.方法 将研究对象分成实验组和对照组,实验组为经肾活检确诊的10例急性排斥反应和12例慢性排斥反应患者,对照组为12例移植肾功能稳定受者和13例健康志愿者.通过ClinProt磁珠浓缩和基质辅助激光解吸电离法(MALDI)-TOF-MS分析两组研究对象的血清蛋白质谱的差异.结果 通过与健康对照组比较,筛选出18条差异性多肽作为诊断肾移植急性排斥反应的潜在生物标记物,6条差异性多肽作为诊断慢性排斥反应的潜在生物标记物.此外,还比较了急性排斥反应与慢性排斥反应患者的多肽差异,检测出了4条高表达的差异性多肽.运用快速分类算法,建立了移植排斥反应的分类模型,该模型对急性排斥反应患者的有效识别能力达到82.64%,对慢性排斥反应患者的有效识别能力高达98.96%.结论 基于功能性磁珠的样本分离法结合MALDI-TOF-MS分析的实验方法 是精确而稳定的.应用蛋白质组学技术建立了肾移植排斥反应患者的血清蛋白质谱诊断模型,为人们更好的理解肾移植排斥反应的发病机制,并将蛋白质组学技术用于排斥反应早期诊断的新技术手段提供了思路.

Variabilities of serum peptidomics spectra in patients with renal allograft rejection

Objective To study the variabilities of serum peptidomics spectra in patients with renal allograft rejection in order to detect specific peptide biomarkers that can be used in the early diagnosis of renal allograft rejection. Methods The serum peptide profiling of two groups of patients with renal allograft rejection (including 10 cases of biopsy-proven acute rejection and 12 cases of chronic rejection) and two groups of controls (including 12 stable graft function transplant recipients and 13 healthy volunteers) was analyzed by the ClinProt magnetic bead enrichment and MALDI-TOF MS. Results A total of 18 differential peptide peaks were selected as potential biomarkers of acute rejection patients,and a total of 6 differential peptide peaks as potential biomarkers of patients with chronic rejection. Between the acute and chronic rejection groups, 4 high expressed differential peptide peaks were detected. A Quick Classifier Algorithm was used to set up the classification model between acute rejection and normal controls, and between chronic rejection and normal controls. The model separated acute rejection from normal controls with a recognition capability of 82.64%,and separated effectively chronic rejection from normal controls with a recognition capability of 98. 96 %. Conclusion Functional magnetic bead based sample fractionation method combined with MALDI-TOF-MS is accurate and stable. This study established a serum peptide fingerprint model for diagnosis of renal allograft rejection.