糖尿病慢性肾脏疾病血清蛋白质组学分析

Serum proteomics analysis in patients with chronic kidney disease in diabetes

目的应用血清蛋白质组学探索糖尿病慢性肾脏疾病(CKD)致病因子和早期诊断指标。方法将研究对象分为正常对照组(NC)、尿白蛋白/肌酐(UACR)<30 mg/g组、30≤UACR<300 mg/g组及UACR≥300 mg/g组。收集各组空腹血清,去除白蛋白,双向电泳分离,银染。分析差异表达蛋白质点,挖取、酶解后进行质谱分析,与Swiss Prot数据库匹配鉴定。结果各组匹配蛋白点共有2119个,挖取11个差异表达蛋白质点进行质谱分析,成功鉴定其中4个蛋白质点。甘露糖结合凝集素相关丝氨酸蛋白酶2(MASP2)在UACR<30 mg/g组表达增高;前凝血酶原在30≤UACR<300 mg/g组表达增高;血红蛋白α亚单位、免疫球蛋白α1(Igα1)链C区段在UACR≥300 mg/g组表达增高。结论临床血清蛋白质组学可作为CKD诊断有效的研究手段。MASP2、前凝血酶原、血红蛋白α亚单位、Igα1链C区段与CKD的发生发展有关。

Objective To explore pathogenic factors for chronic kidney disease （CKD）and to investigate biomarkers of early diagnosis of CKD by serum proteomics analysis. Methods Study subjects enrolled in this study were divided into normal control group（NC）, urinary albumin/creatinine（UACR）less than 30 mg/g group, 30 to 300 mg/g group, and more than 300 mg/g group. Fasting plasma was collected. After detaching albumin, all the plasma with equivalent protein was separated with two- dimensional electrophoresis（2DE）, and then stained by silver staining kit. The proteins that expressed significantly different among the four groups were determined and detected with mass spectrometry（MS）. Finally the proteins were matched and Certificated in Swiss Prot Bank. Results A total of 2119 protein spots were detected and matched after separation with 2DE among each group. Eleven spots were dug and four of them were successfully determined with MS. The expression of mannan-binding lectin serine protease 2 （MASP2） elevated significantly in less than 30 mg/g group. Prothrombin expressed much higher in 30 to 300 mg/g group. Meanwhile, the expression of hemoglobin subunit alpha and Ig alpha-1 chain C region increased significantly in more than 300 mg/g group. Conclusion Clinical serum proteomics is one of valuable and efficient tool in clinical practice and research of CKD. MASP2, prothrombin, hemoglobin subunit alpha, and Ig alpha-1 chain C might be associated with the development of CKD.