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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/ionizationtime 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5.5mg/kg复制MCNS模型,造模10d后分别予以强的松和蒸馏水灌胃治疗。21d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。
Objective To investigate the expression of minimal-changed nephrotic syndrome (MCNS) using surface enhanced laser desorption / ionizationtime of flight-mass spectrometry (SELDI-TOF-MS) Significance of Screening Urine Biomarkers in Rats. Methods Thirty SD rats were randomly divided into normal control group (n = 10), model control group (n = 10) and prednisone treatment group (n = 10). Except for the normal control group, the other rats were injected with doxorubicin 5.5 mg / kg through the tail vein to establish a model of MCNS. After 10 days of modeling, prednisone and distilled water were given to the rats respectively. Urine was collected after 21 days, and protein was captured by weak cation exchange chip CM10. Urinary protein fingerprints of three groups of rats were obtained by SELDI-TOF-MS. Biomarker software was used to analyze differential proteins. Results The optimal urine protein fingerprint was obtained after optimizing the experimental conditions. The average CV value of the control peak intensity was 18.0% (7.50% -35.89%). There were 33 statistically significant differences in protein peaks (P <0.05) between the model control group and the normal control group, of which 21 were up-regulated and 12 were down-regulated. There was a significant difference in protein expression between the model control group and prednisone treatment group (P <0.05) in the normal control group. These differential protein peaks were highly expressed in the model control group, while in the prednisone treatment group, protein expression Down, peak intensity close to the normal control group. Conclusion SELDI-TOF-MS can provide a feasible method for the screening of urine biomarkers in MCNS rats by optimizing the experimental conditions and implementing quality control. The differential protein peaks obtained are of great significance for the early diagnosis of MCNS, the monitoring of treatment and the search for target of drug therapy.