Journal
CLINICAL BIOCHEMISTRY
Volume 45, Issue 18, Pages 1613-1619Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.clinbiochem.2012.08.027
Keywords
Zymography-LC-MS/MS; Nephropathy; Retinopathy; Saliva; Urine
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Funding
- Portuguese Foundation for Science and Technology (FCT) [SFRH/BD/30124/2006, PTDC/QUI/72683/2006, PEst-C/QUI/UI0062/2011]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/30124/2006, PTDC/QUI/72683/2006] Funding Source: FCT
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Objectives: We aimed to disclose the proteolytic events underlying type 1 diabetes and related complication through protease profiling in the bodily fluids serum, urine and saliva. Design and methods: Zymography followed by LC-MS/MS was performed for protease identification and quantitative comparison of proteolytic activity between healthy, type 1 diabetic patients with no complications and with retinopathy and nephropathy. Western blotting was also accomplished for MMP-9 and MMP-2 identification and expression analysis. Results: Only MMP-2 and MMP-9 were observed in serum with significantly increased levels and activity observed in diabetic patients. In urine and saliva other proteases besides MMPs were identified by MS and presented disease-dependent activity variations. Among these are complex MMP-9/Neutrophil gelatinase-associated lipocalin, aminopeptidase N, azurocidin and kallikrein 1 with more activity noticed in type 1 diabetes patients with nephropathy and/or retinopathy. Conclusion: Our data highlight the usefulness of urine and saliva for the monitoring of type-1 diabetes-related proteolytic events, where aminopeptidase N, azurocidin and kallikrein 1 appear as promising screening targets for type 1 diabetes-related complications. (C) 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
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