Ⅲ～Ⅳ期糖尿病肾病气阴两虚证患者的血清蛋白质组学研究

Serum proteomics study on Ⅲ-Ⅳ stage of diabetic nephropathy with Qi-Yin deficiency syndrome

目的应用表面加强激光解吸电离-飞行时间-质谱(SELDI-TOF-MS)技术,从蛋白质组学角度筛选Ⅲ～Ⅳ期糖尿病肾病(DN)患者气阴两虚证的差异蛋白。方法采集37例Ⅲ～Ⅳ期气阴两虚证DN患者(DN气阴两虚证组)、33例Ⅲ～Ⅳ期非气阴两虚证DN患者(DN非气阴两虚证组)和35例健康受试者(健康对照组)的血清样本,采用SELDI-TOF-MS技术检测各组血清蛋白表达谱,后运用Biomarker WizardTM和Biomarker PatternsTM软件进行数据分析,筛选出特异蛋白,建立诊断决策树模型。结果 DN气阴两虚证组与健康对照组共有8个蛋白峰比较差异有统计学意义(P均<0. 05); DN气阴两虚证组与DN非气阴两虚证组共有11个蛋白峰比较差异有统计学意义(P均<0. 05);质荷比为4 986. 15Da所代表的蛋白可能在Ⅲ～Ⅳ期DN气阴两虚证患者中呈特异表达;以质荷比为1 084. 57 Da等11个蛋白组成的证候决策树模型,敏感性为97. 3%,特异性为95. 6%。结论Ⅲ～Ⅳ期DN气阴两虚证的发生发展,可能是以质荷比为4 986. 15 Da蛋白的差异表达为物质基础的。

Objective It is to apply the technology of surface-enhanced laser desorption/ionization-time of flight-mass spectrometry(SELDI-TOF-MS)to detect the specific serum protein for Ⅲ-Ⅳ stage of diabetic nephropathy(DN)with Qi-Yin deficiency syndrome.Methods Seventy serum samples from Ⅲ-Ⅳ stage of DN patients(37 samples with Qi-Yin deficiency syndrome,33 samples with non-Qi-Yin deficiency syndrome)and 35 serum samples from healthy persons were collected.SELDI-TOF-MS technology was used to detect the expression profile of serum protein,and then use the softwares of Biomarker WizardTM and Biomarker WizardTM to make data analysis.In the end,the special serum protein can be identified,and the decision model was established.Results Eight difference protein peaks can be distinction between DN patients of Qi-Yin deficiency syndrome group and healthy group.Eleven difference protein peaks can be distinction between Qi-Yin deficiency syndrome group and non-Qi-Yin deficiency syndrome group in DN patients(P<0.05).Combined to the above mentions and statistical analysis,the protein of M/Z 4 986.15 was likely to be the serum protein markers for Ⅲ-Ⅳ stage of DN with Qi-Yin Deficiency Syndrome.A decision model consisted by 11 protein peaks(just like M/Z 1 084.57 and so on)can be built,with sensitivity of 97.3%,specificity of 95.6%.Conclusion The protein of M/Z 4986.15 is the material foundation for development about Ⅲ-Ⅳ stage of DN with Qi-Yin DN.