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J. Yu, F. Wang; The Differential Vitreous Proteome of Proliferative Vitreoretinopathy. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6091.
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© ARVO (1962-2015); The Authors (2016-present)
To fish the special vitreous protein in rhegmatogenous retinal detachment patients with proliferative vitreoretinopathy (PVR), vitreous proteomes were investigated by two-dimensional gel electrophoresis coupled with mass spectrometry.
Vitreous samples of moderate PVR (grade B), and severe PVR (grade C or D) were aspirated during pars plana vitrectomy (PPV) before infusion were collected preoperatively. The normal vitreous from donor eyes was as the control. First dimension isoelectric focusing (IEF) was performed with the Pharmacia IPGphor in solvent B using 18 cm non-linear pH 3-10 immobilized pH gradient (IPG) strips. IPG strips were rehydrated with sample before IEF was performed. The second dimension 12% sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) was performed after the IPG strips were equilibrated. After silver staining, the gels were analyzed by the 2-DE gel analysis software. The matched spots were excised and tryptic digested in-gel. The pepetides were analyzed by MALDI-TOF-MS and the MS/MS spectras were searched against the human protein databases using MASCOT.
In the current study, there were 47, 184 and 336 protein spots were analyzed from the 2-DE gels of donor, moderate PVR and severe PVR. Among 13 the significantly difference spots, 7 proteins were successfully identified. Enolase 2 was the special protein in donor vitreous; transthyretin monomer and retinol-binding protein (RBP) chain B were special proteins in severe PVR vitreous; prostaglandin D synthase and RBP3 precursor were overlaped,which were upregulated in moderate PVR but downregulated in severe PVR; albumin and transferrin were upregulated associated with the severity of PVR.
Our current proteome study presented that normal proteins reduced and some common serum proteins upregulated, which indicated that blood-retinal barrier was destroyed in the process of PVR.
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