基于表面增强激光解析离子化飞行时间质谱技术的慢性肾衰竭中医湿证尿液中相关蛋白研究

Study on Correlated Proteins in the Urine of Chronic Renal Failure Patients of Chinese Medicine Damp Syndrome Based on SELDI-TOF-MS Technique

目的采用表面增强激光解析离子化飞行时间质谱(surface-enhanced laser desorption/ionization time of flight mass spectrometry,SELDI-TOF-MS)技术研究慢性肾衰竭(chronic renal failure,CRF)中医湿证患者的尿液蛋白标志物。方法采集CRF湿证患者90例和非湿证患者60例的尿液,采用H4蛋白芯片技术进行尿液蛋白质组学研究,用蛋白芯片阅读器PBSⅡ对芯片进行扫描、分析。结果 (1)湿证组与非湿证组尿液样本的蛋白质图谱在质荷比1000-20000范围内检测到25个差异蛋白峰(P<0.01)。(2)经生物信息学分析建立CRF中医湿证尿液蛋白预测模型,得到M/Z8654.96、M/Z2081.65、M/Z18667.3和M/Z2242.14共4个差异蛋白峰组成的生物标记物可以将湿证组和非湿证组样本较好地分类,其正确率84.7%,灵敏度为92.2%,特异性为73.3%。(3)湿证组与非湿证组尿液中差异蛋白峰经SwissProt数据库鉴定,可能为7种蛋白质。结论初步筛选出CRF中医湿证的尿液蛋白标志物,建立了CRF中医湿证尿液蛋白预测模型,通过数据库对尿液蛋白标志物进行了鉴定,为CRF中医湿证的临床辨证提供了一定的实验依据。

Objective To investigate the protein markers in the urine of chronic renal failure （CRF） patients of Chinese medicine damp syndrome （CMDS） based on surface-enhanced laser desorption/ionization time of flight mass spectrometry （SELDI-TOF-MS） technique. Methods The urine was sampled from 90 CRF patients of CMDS and 60 CRF patients of non-CMDS. Then the proteomics of the urine was studied by H4 gene chip. The chips were scanned and analyzed using PBSⅡ （a protein chip reader）. Results （1） Totally 25 differential protein peaks were identified in the e/m range of 1 000-20 000 of the protein atlas of the two groups （P0.01）. （2） The urine protein predictive model of CFR patients of CMDS was established after bioinformatic analysis. Totally 4 biomarkers consisting of M/Z 8 654.96, M/Z 2 081.65, M/Z 18 667.3, and M/Z 2 242.14 were obtained, which could better classify the samples of CMDS and those of non-CMDS. Its accuracy rate was 84.7%, the sensitivity was 92.2%, and the specificity was 73.3%. （3） Between the CMDS group and the non-CMDS group, 7 kinds of proteins in the urine were possibly identified by SwissProt Database. Conclusions This study had primarily screened the protein markers in the urine of CRF patients of CMDS, and established the predictive model. The protein markers in the urine were identified by database, thus providing certain experimental evidence for clinical typing of CRF patients of CMDS.