SELDI-TOF-MS筛选微小病变型肾病综合征尿液生物标志物.docVIP

SELDI-TOF-MS筛选微小病变型肾病综合征尿液生物标志物 林 青1，郑 健2，陈蓉艳1，艾 斯2 （350004福建 福州，福建中医学院附属人民医院：1检验科，2儿科） [摘要]目的 探讨应用优化的表面增强激光解析电离-飞行时间-质谱（surface enhanced laser desorption/ionization time of flight-mass spectrometry ，SELDI-TOF-MS）进行微小病变型肾病综合征（minimal-changed nephrotic syndrome，MCNS）大鼠尿液生物标志物筛选的意义。方法 SD大鼠30只随机分为正常对照组、模型对照组和强的松治疗组。除正常对照组外，其余大鼠经尾静脉一次性注射阿霉素5.5 mg/kg复制MCNS模型，造模10 d后分别予以强的松和蒸馏水灌胃治疗。21 d后收集大鼠尿液，经弱阳离子交换芯片CM10捕获蛋白质，应用SELDI-TOF-MS获取三组大鼠的尿液蛋白质指纹图谱，采用Biomarker wizard软件比对分析差异蛋白。结果 优化实验条件后获得较理想的尿液蛋白质指纹图谱，质控峰强度的平均CV值为18.0 %（7.50 %～35.89 %）。在模型对照组与正常对照组之间检测到不同质荷比处有33个具统计学意义的差异蛋白峰（P 0.05），其中21个蛋白表达上调，12个蛋白表达下调。在正常对照组、模型对照组与强的松治疗组之间有六个蛋白质表达存在差异（P 0.05），这些差异蛋白峰在模型对照组中高表达，而在强的松治疗组中蛋白表达下调，峰强度接近正常对照组。结论 通过优化实验条件并实行质量控制，SELDI-TOF-MS可为MCNS大鼠尿液生物标志物的筛选提供一个可行的检测途径。所获得的差异性蛋白峰对MCNS的早期诊断、治疗监测和寻找药物治疗的靶目标具有重要的意义。 [关键词]SELDI-TOF-MS；微小病变型肾病综合征；尿液；生物标志物；质控 [中图法分类号] [文献标志码]A SELDI-TOF-MS as a method for biomarker discovery in the urine of minimal-changed nephrotic syndrome LIN Qing1, ZHENG Jian2, CHEN Rong-yan1, AI Si2 (1Department of Clinical Laboratory, 2Department of Paediatrics, The Affiliated People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou 350004,China) [Abstract] Objective To study the use of optimal surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) for detecting biomarkers in the urine of minimal-changed nephrotic syndrome(MCNS). Methods Thirty SD rats were randomly divided into normal control group, untreated group and prednisone group. Except the normal control group, rats were injected once via caudal vein with adriamycin ( 5.5 mg/kg ) to induce MCNS. 10 days later, the rats were administered with prednisone or distilled water for 21d , respetcively. Collecting the urine of rats, urinary peptide or protein peaks were detected by SELDI-TOF-MS and weak cation exchange proteinchip CM10. The differentially expressed protein and polypeptides were analyzed with Biomarker Wizard Software