PURPOSE: To identify differential changes in proteins and metabolites underlying "fast" type 1 (T1DC) and "slow" type 2 (T2DC) diabetic cataract (DC) formation in rat. METHODS: Rat models of types 1 and 2 diabetes consisted of streptozotocin injection without and with high-fat diet, respectively. Cataract progression was examined weekly. At week 6, total protein changes were comparatively and quantitatively assessed by two-dimensional differential in-gel electrophoresis (2-D DIGE) coupled with mass spectrometry, and relevant metabolic changes were examined. Differences in high molecular weight (HMW) crystallin species between diabetic and control lenses were similarly identified. RESULTS: Cataracts were morphologically different and progressed more slowly in T2DC versus T1DC. αA-crystallin, βB2-crystallin, and βA4-crystallin were significantly decreased in both DC types versus control. αB-crystallin was increased while βB1-crystallin was markedly decreased in T2DC. In T1DC, γB-crystallin and γS-crystallin fragmentation were increased. High-fat diet by itself had little impact, except for lowering γS-crystallin fragmentation. Despite significantly decreased opacity, a greater decrease in intermediate filaments (IFs) and more HMW crystallin species were observed in T2DC versus T1DC. However, aldose reductase expression and activity and sorbitol levels were increased to a greater extent in T1DC, while reduced glutathione (GSH) and reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) levels were decreased to a greater extent and adenosine triphosphate (ATP) level was much lower in T1DC versus T2DC. CONCLUSIONS: The results suggest that osmotic damage, GSH loss, and decreased ATP production might be important pathological mechanisms in T1DC formation, whereas crystallin modification and cross-linking/aggregation as well as IF degradation may play more crucial roles in T2DC formation. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: To identify differential changes in proteins and metabolites underlying "fast" type 1 (T1DC) and "slow" type 2 (T2DC) diabetic cataract (DC) formation in rat. METHODS:Rat models of types 1 and 2 diabetes consisted of streptozotocin injection without and with high-fat diet, respectively. Cataract progression was examined weekly. At week 6, total protein changes were comparatively and quantitatively assessed by two-dimensional differential in-gel electrophoresis (2-D DIGE) coupled with mass spectrometry, and relevant metabolic changes were examined. Differences in high molecular weight (HMW) crystallin species between diabetic and control lenses were similarly identified. RESULTS:Cataracts were morphologically different and progressed more slowly in T2DC versus T1DC. αA-crystallin, βB2-crystallin, and βA4-crystallin were significantly decreased in both DC types versus control. αB-crystallin was increased while βB1-crystallin was markedly decreased in T2DC. In T1DC, γB-crystallin and γS-crystallin fragmentation were increased. High-fat diet by itself had little impact, except for lowering γS-crystallin fragmentation. Despite significantly decreased opacity, a greater decrease in intermediate filaments (IFs) and more HMW crystallin species were observed in T2DC versus T1DC. However, aldose reductase expression and activity and sorbitol levels were increased to a greater extent in T1DC, while reduced glutathione (GSH) and reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) levels were decreased to a greater extent and adenosine triphosphate (ATP) level was much lower in T1DC versus T2DC. CONCLUSIONS: The results suggest that osmotic damage, GSH loss, and decreased ATP production might be important pathological mechanisms in T1DC formation, whereas crystallin modification and cross-linking/aggregation as well as IF degradation may play more crucial roles in T2DC formation. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
Entities:
Keywords:
2-D DIGE; diabetic cataract; lens proteins; mass spectrometry; proteomics
Authors: Jeremy A Whitson; Phillip A Wilmarth; John Klimek; Vincent M Monnier; Larry David; Xingjun Fan Journal: Free Radic Biol Med Date: 2017-09-22 Impact factor: 7.376