ClinProt技术在子宫内膜异位症研究中的应用

【作者】 王琳；

【导师】 郎景和；

【作者基本信息】 北京协和医学院 ， 临床医学， 2010， 博士

【摘要】 子宫内膜异位症(内异症)是常见的妇科疾患,发病率在育龄期妇女约10-15%,且有逐渐上升的趋势。该疾病的主要表现为慢性盆腔痛、痛经、不孕和附件包块,严重的影响了妇女的生活质量。同时虽然子宫内膜异位症是一种良性疾病,但具有侵袭、转移与复发等恶性行为。研究表明,早期的内异症病变主要集中在腹膜,对盆腔结构的影响不大,早期诊断和治疗有助于患者生活质量的提高。目前,关于内异症的早期诊断,尚且没有有效的标志物,没有标志物广泛应用于临床,用以评价疾病的严重程度判断、评估疗效和术后随访。本实验采用ClinProt技术联合MALDI-TOF,建立子宫内膜异位症患者腹腔液在分子量800-10000Da范围内的蛋白质/肽图谱,期望寻找该病的特异性蛋白,并鉴定蛋白性质,明确该蛋白在内异症中的作用,为早期诊断和发病机制的研究提供依据。同时使用ClinProt技术联合MALDI-TOF技术建立内异症患者尿中的低分子量蛋白质/肽图谱,建立高效率诊断模型。该实验通过联合MALDI-TOF/TOF和LC-MS/MS,鉴定出差异蛋白为胶原的前体,可能在内异症的发生发展过程中起到重要作用。本实验基于ClinProt技术建立的诊断模型效率高,将来该技术有可能广泛应用于临床,实现对该病的的无创诊断,为内异症的早期诊断和治疗随访提供依据,同时也为将来子宫内膜异位患者的腹腔液和尿中的蛋白质组学研究提供了经验和数据。更多还原

【Abstract】 OBJECTIVESThis study aimed to to investigate the possibility of using the ClinProt technique to find diagnostic markers that are able to better discriminate healthy subjects from patients affected by endometriosis(EMs). Endometriosis is a common gynecological disease. Biomakers for early detection, prognosis, follow-up, and differential diagnosis of Ems are needed in daily clinical practice.METHODSPeritoneal fluid and urine samples were analyzed by the ClinProt and MALDI-ToF technique. We combined the automated high through put MALDI-TOF measurements and LC-MS/MS to identify the different proteins.RESULTSAfter comparing the peritoneal fluid protein mass spectra of39patients with34controls,18differently expressed potential biomarkers (P<0.01) were identified. Using ClinproTool software, we established GA algorithm.When using the a cluster of five signals (m/z=4458.23,2379.3,8519.94,6376.99,6190.27) to classify the samples, the sensitivity for diagnosing endometriosis was88.5%, specificity was88.5%.Urine samples from30patients and31controls were also analyszed by MALDI-TOF. Using GA algorithm, The combination of five markers (m/z=1433.98,1599.48,2085.61,6798.07,3217.29) lead to an improvement in diagnostic efficacy, with specificity and sensitivity of95.7%and96.7%, respectively.Com bind ing the MALDI-TOF/TOF with LC-MS/MS, we identified the proteins as collagen precursor.CONCLUSIONSThese results may be useful to set up new diagnostic or prognostic tools.更多还原