液体芯片-飞行时间质谱对结膜松弛症患者泪液蛋白质的分析

Analysis of tear proteins from conjunctivochalasis patients by liquid chip time-of-flight mass spectrometry

目的应用液体芯片-飞行时间质谱系统分析结膜松弛症患者的泪液蛋白质表达谱,寻找潜在具有结膜松弛症诊断意义的生物学标志物。方法收集10例结膜松弛症患者和9例正常对照的泪液,利用毛细管法每眼收集15μL泪液。经ClinProt磁珠纯化、基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析及ClinProTools生物信息学软件研究其泪液蛋白表达谱,比较2组人群泪液蛋白质谱表达的差异。结果 2组共检测发现了77个蛋白质峰,其中13个蛋白质峰差异有统计学意义(P<0.05)。其中质荷比(m/z)为1 632.48和3 401.87者为差异蛋白质,能较好地鉴别结膜松弛症组和正常对照组。结论液体芯片-飞行时间质谱系统可高效、精确地筛查泪液蛋白质。结膜松弛症组与正常对照组泪液蛋白存在差异,值得进一步研究,可能对发现结膜松弛症的发病机制和寻求有效的防治手段有指导作用。

Background Tear is a complex biological mixture.Conjunctivochalasis causes the abnormality of tear function and ocular surface.The analysis of tear proteomics changes in conjunctivochalasis helps to identify the pathogenesis of conjunctivochalasis.ObjectiveThis study was to analyze the expression profile of tear proteins in patients with conjunctivochalasis by liquid chip time-of-flight mass spectrometry and search for potential diagnostic value of biomarkers for conjunctivochalasis.MethodsTears were collected from 10 eyes of 10 conjunctivochalasis patients and 9 eyes of 9 matched normal volunteer,and 15 μL of tears were obtained by microcapillary tubes from each eye.The tear samples were purified by the ClinProt magnetic bead enrichment and analyzed by the MALDI-TOF-MS.The expression mass spectra of tear protein was analyzed by ClinProTools bioinformation software to compare the difference between conjunctivochalasis patients and normal subjects.Written informed consent was obtained from all of the individuals before examination.ResultsSeventy-seven protein peaks were detected in human tear with the relative molecular wight from 0-40 000.The 13 significant different expressive peaks were screeded from 77 kinds of proteins in tear between conjunctivochalasis group and normal control group（P0.05）.Two kinds of proteins with the mass charge ratio（m/z） of 1632.48 and 3401.87 showed the lower protein abundance in human tear of conjunctivochalasis group than the normal control group（P=0.025,P=0.025）,being optimal differentiatial proteins between conjunctivochalasis and normal control group.ConclusionThe liquid chip time-of-flight mass spectrometry system can achieve efficient,accurate screening of tear protein.Differential protein profile change is found in the tear of conjunctivochalasis patient.Further study should be performed to explore the pathogenesis mechanism and seek effective prevention of conjunctivochalasis.