Proteomic analysis of human vitreous fluid by fluorescence-based difference gel electrophoresis (DIGE): a new strategy for identifying potential candidates in the pathogenesis of proliferative diabetic retinopathy

Garcia-Ramírez, Marta; Canals, Françesc; Hernández, Cristina; Colomé, Núria; Ferrer, Cristina Suárez; Carrasco, Esther; García‐Arumí, José; Simó, Rafael

doi:10.1007/s00125-007-0627-y

Cited by 151 publications

(150 citation statements)

References 36 publications

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“…Some of the identified proteins appeared at multiple positions on the gels in this and other studies [6,16], which is consistent with the presence of different post-translationally modified forms. Post-translational modifications can change the MW and the pI of proteins, and the various forms of these proteins can migrate to different spots.…”

Section: Discussionsupporting

confidence: 90%

“…PEDF alterations in patients with PDR compared with nondiabetic patients are controversial. Some previous studies pointed to reductions in the levels of vitreous PEDF in patients suffering from PDR [6,18-20]. Conversely, elevated levels of PEDF were detected in some studies [13,21].…”

Section: Discussionmentioning

confidence: 95%

“…Currently, modern proteomic technologies have the advantage of facilitating the simultaneous analysis and identification of large numbers of proteins. Recent advances in proteome analysis methods have allowed the further exploration and acquisition of vitreous protein profiles [1,6-8]. In the last few years, proteomics has been applied to explore proteins which were differentially expressed in patients with proliferative diabetic retinopathy and in nondiabetic patients [9-14].…”

Section: Introductionmentioning

confidence: 99%

“…The technique of two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) provides an accurate quantitative comparison of two groups of samples, allowing the identification of proteins whose levels differ significantly between the two conditions [6]. No previous quantitative proteomic comparison of vitreous humour from type 2 diabetic patients with PDR with that from normal human eyes donated for corneal transplant has been reported.…”

Section: Introductionmentioning

confidence: 99%

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Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Wang

Hu³

et al. 2012

Proteome Sci

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BackgroundDiabetes can lead to serious microvascular complications such as proliferative diabetic retinopathy (PDR), which results in severe vision loss. The diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. The vitreous humour can be utilised in a variety of studies aimed toward the discovery of new targets for the treatment or prevention of PDR and the identification of novel disease mechanisms. The aim of this study was to compare the protein profile of vitreous humour from diabetic patients with PDR with that of vitreous humour from normal human eyes donated for corneal transplant.ResultsVitreous humour from type 2 diabetic patients with PDR (n = 10) and from normal human eyes donated for corneal transplant (n = 10) were studied. The comparative proteomic analysis was performed using two-dimensional fluorescence difference gel electrophoresis (2-D DIGE). Differentially produced proteins (abundance ratio > 2 or < -2, p < 0.01) were identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF tandem mass spectrometry. A total of 1242 protein spots were detected on the 2-D master gel of the samples, and 57 spots that exhibited statistically significant variations were successfully identified. The spots corresponded to peptide fragments of 29 proteins, including 8 proteins that increased and 21 proteins that decreased in PDR. Excluding the serum proteins from minor vitreous haemorrhage, 19 proteins were found to be differentially produced in PDR patients compared with normal subjects; 6 of these proteins have never been reported to be differentially expressed in PDR vitreous: N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 (DDAH 1), tubulin alpha-1B chain, gamma-enolase, cytosolic acyl coenzyme A thioester hydrolase, malate dehydrogenase and phosphatidylethanolamine-binding protein 1 (PEBP 1). The differential production of pigment epithelium-derived factor (PEDF) and clusterin was confirmed by Western blot analysis.ConclusionsThese data provide an in-depth analysis of the human vitreous proteome and reveal protein alterations that are possibly involved in the pathogenesis of PDR. Further investigation of these special proteins may provide potential new targets for the treatment or the prevention of PDR.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Wang

Hu³

et al. 2012

Proteome Sci

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“…We recently compared the protein profiles of human vitreous from diabetic patients with proliferative diabetic retinopathy (PDR) with the vitreous fluid from non-diabetic patients with macular holes using the fluorescence-based difference gel electrophoresis (DIGE) strategy [15]. Our results provided evidence that IRBP is among the lowest produced proteins in the vitreous fluid of patients with PDR.…”

Section: Introductionmentioning

confidence: 99%

Interphotoreceptor retinoid-binding protein (IRBP) is downregulated at early stages of diabetic retinopathy

et al. 2009

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Aims/hypothesis Interphotoreceptor retinoid-binding protein (IRBP) plays a major role in the visual cycle and is essential to the maintenance of photoreceptors. The aim of this study was to determine whether a decrease in IRBP production exists in the early stages of diabetic retinopathy. Methods Vitreous samples from diabetic patients with proliferative and non-proliferative diabetic retinopathy (PDR, NPDR), and from non-diabetic patients with macular hole (control group) were selected for IRBP quantitative assessment by proteomic analysis (fluorescence-based difference gel electrophoresis) and western blot. Human post mortem eyes (n=16) from diabetic donors without clinically detectable retinopathy and from non-diabetic donors (n=16) were used to determine IRBP (also known as RBP3) mRNA levels (RT-PCR) and protein content (western blot and confocal microscopy). Retinal neurodegeneration was assessed by measuring glial fibrillar acidic protein (GFAP) and the apoptotic rate. Y79 human retinoblastoma cells were used to test the effects of glucose, TNF-α and IL-1β on IRBP expression and IRBP levels. Results Intravitreous IRBP concentration was significantly lower in PDR

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Elevation of Apolipoprotein A-I and Apolipoprotein H Levels in the Vitreous Fluid and Overexpression in the Retina of Diabetic Patients

Simó

Higuera²,

Garcia-Ramírez³

et al. 2008

Arch Ophthalmol

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Objectives: To determine levels of apolipoprotein (apo) A-I and apo H in the vitreous fluid of patients with proliferative diabetic retinopathy (PDR) and to examine whether apo A-I and apo H messenger RNA (mRNA) levels are overexpressed in the diabetic retina. Methods: Vitreous samples from 4 diabetic patients with PDR and 8 nondiabetic patients with macular hole were selected for proteomic analysis. Fourteen additional samples (7 from patients with PDR and 7 from patients with macular hole) were used for Western blot analysis. Fourteen postmortem eyes (7 from diabetic and 7 from nondiabetic donors) were used to perform quantitative real-time polymerase chain reaction analysis. Results: Intravitreous apo A-I and apo H levels were significantly higher in patients with PDR than in the control group. The apo A-I and apo H mRNA levels obtained from the retinas of diabetic donors were significantly higher than those obtained from nondiabetic donors. Retinal pigment epithelium was the main contributor to the differences. Conclusions: Levels of apo A-I and apo H are elevated in the vitreous fluid of diabetic patients with PDR. In addition, we provide the first evidence, to our knowledge, that a higher expression of apo A-I and apo H mRNAs exists in the diabetic retina. Clinical Relevance: The results of this study may be relevant to new treatment strategies aimed toward reducing the development of diabetic retinopathy.

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Proteomic analysis of human vitreous fluid by fluorescence-based difference gel electrophoresis (DIGE): a new strategy for identifying potential candidates in the pathogenesis of proliferative diabetic retinopathy

Cited by 151 publications

References 36 publications

Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Interphotoreceptor retinoid-binding protein (IRBP) is downregulated at early stages of diabetic retinopathy

Elevation of Apolipoprotein A-I and Apolipoprotein H Levels in the Vitreous Fluid and Overexpression in the Retina of Diabetic Patients

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