SELDI-TOF-MS筛选微小病变型肾病综合征尿液生物标志物

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(n=10)、模型对照组(n=10)和强的松治疗组(n=10).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3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6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.

Screening of biomarkers in urine of rats with minimally-changed nephrotic syndrome by surface enhanced laser desorption/ionization time of flight-mass spectrometry

Objective To detect the biomarkers in urine of rats with minimally-changed nephrotic syndrome (MCNS) by optimal surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS). Methods Thirty SD rats were randomly divided into normal control group,untreated group and prednisone treatment group. Except for the normal control group,rats were injected with adriamycin (5.5 mg/kg once) via caudal vein to induce MCNS. Ten days later,the rats were administered with prednisone or distil...