Liu Moyan, Yu Hanjie, Zhang Dong, Han Qiuxia, Yang Xiaoli, Liu Xiawei, Wang Jifeng, Zhang Kun, Yang Fuquan, Cai Guangyan, Chen Xiangmei, Zhu Hanyu
Department of Nephrology, Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center of Kidney Diseases 28 Fuxing Road, Haidian District Beijing 100853 China
Second Department of Cadre Ward, General Hospital of Jinan Military Region Jinan 250000 China.
RSC Adv. 2018 Nov 19;8(68):38872-38882. doi: 10.1039/c8ra06832a. eCollection 2018 Nov 16.
Diabetic nephropathy (DN) and nondiabetic renal disease (NDRD) are two major categories of renal diseases in diabetes mellitus patients. The clinical differentiation among them is usually not so clear and effective. In this study, sera from DN and NDRD patients were collected, and glycan profiles of serum proteins from DN and NDRD patients were investigated and compared by using lectin microarray and lectin blot. Then, altered glycoproteins were enriched by lectin coupled magnetic particle conjugate and characterized by LC-MS/MS. We found significant change in glycan patterns between DN and NDRD patients. In particular, the relative abundance of the glycopattern of Galβ1-3GalNAc which was identified by BPL ( lectin) was significantly decreased in DN patients compared to four types of NDRD patients ( < 0.05). Moreover, BPL blotting indicated that the proteins with a molecular weight of about 53 kDa exhibited low staining signal in DN compared to all NDRD groups, which was consistent with results of lectin microarrays. After enriching by BPL and identification by LC-MS/MS, a total of 235 and 258 proteins were characterized from NDRD and DN respectively. Among these, the relative abundance of 12 isolated serum proteins exhibited significantly alteration between DN and NDRD ( < 0.05). Our findings indicated not only the relative abundance of Galβ1-3GalNAc on serum proteins but also certain glycoproteins modified with this glycopattern showed a difference between DN and NDRD patients. This suggested that the analysis of this alteration by using urine specimens may constitute an additional valuable diagnostic tool for differentiating DN and NDRD with a non-invasive method.
糖尿病肾病(DN)和非糖尿病性肾脏疾病(NDRD)是糖尿病患者中两类主要的肾脏疾病。它们之间的临床鉴别通常并不清晰有效。在本研究中,收集了DN和NDRD患者的血清,采用凝集素芯片和凝集素印迹法对DN和NDRD患者血清蛋白的聚糖谱进行了研究和比较。然后,通过凝集素偶联磁珠共轭物富集改变的糖蛋白,并通过液相色谱-串联质谱(LC-MS/MS)进行表征。我们发现DN和NDRD患者之间的聚糖模式有显著变化。特别是,与四种类型的NDRD患者相比,由BPL(凝集素)鉴定的Galβ1-3GalNAc糖模式的相对丰度在DN患者中显著降低(<0.05)。此外,BPL印迹表明,与所有NDRD组相比,分子量约为53 kDa的蛋白质在DN中显示出低染色信号,这与凝集素芯片的结果一致。通过BPL富集并经LC-MS/MS鉴定后,分别从NDRD和DN中鉴定出235种和258种蛋白质。其中,12种分离的血清蛋白的相对丰度在DN和NDRD之间表现出显著变化(<0.05)。我们的研究结果表明,不仅血清蛋白上Galβ1-3GalNAc的相对丰度,而且某些被这种糖模式修饰的糖蛋白在DN和NDRD患者之间也存在差异。这表明,利用尿液标本分析这种变化可能构成一种额外的有价值的诊断工具,用于以非侵入性方法区分DN和NDRD。