Diabetic nephropathy (DN) is a long-term complication of diabetes mellitus that leads to end-stage renal disease. Microalbuminuria\r\nis used for the early detection of diabetic renal damage, but such levels do not reflect the state of incipient DN precisely in type\r\n2 diabetic patients because microalbuminuria develops in other diseases, necessitating more accurate biomarkers that detect incipient\r\nDN. Isobaric tags for relative and absolute quantification (iTRAQ) were used to identify urinary proteins that were differentially\r\nexcreted in normoalbuminuric and microalbuminuric patients with type 2 diabetes where 710 and 196 proteins were identified\r\nand quantified, respectively. Some candidates were confirmed by 2-DE analysis, or validated by Western blot and multiple\r\nreaction monitoring (MRM). Specifically, some differentially expressed proteins were verified by MRM in urine from normoalbuminuric\r\nand microalbuminuric patients with type 2 diabetes, wherein alpha-1-antitrypsin, alpha-1-acid glycoprotein 1, and prostate\r\nstem cell antigen had excellent AUC values (0.849, 0.873, and 0.825, resp.). Moreover, we performed a multiplex assay using\r\nthese biomarker candidates, resulting in amerged AUC value of 0.921. Although the differentially expressed proteins in this iTRAQ\r\nstudy require further validation in larger and categorized sample groups, they constitute baseline data on preliminary biomarker\r\ncandidates that can be used to discover novel biomarkers for incipient DN.
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