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S.H. Lee, K.W. Park, C.I. Kim, Y.J. Cho, W.I. Yoo, Y.S. You, S.J. Lee, O.W. Kwon; Proteomics Analysis of Diabetic Retinopathy Vitreous Body . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2238.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Development of diabetic retinopathy is characterized by loss of pericyte and increased vascular permeability, blood-retinal barrier breakdown, followed by microvascular occlusion and neovascularization in the retinas. We thought that the diffusible factors and proteins associated with pathogenesis should be present with the vitreous body. We used a proteomics-based approach to reveal vitreous proteom alteration that elicit proliferative diabetic retinopathy(PDR). Methods: Human vitreous samples were obtained from 10 patients with non-insuline dependent diabetes mellitus(NIDDM) at the time of vitrectomy for treatment of PDR. And Cadaveric control vitreous was obtained within 5 to 7hr after death from eye donors who had no known ocular or systemic inflammatory disease. Proteins were separated by 2-Dgel electrophresis(pH 3-10; 8-18% gradient). Protein spots were cut from coomassie blue-stained gels, digest with trypsin, and identified by using a MALDI mass spectrometry. And then some samples were analysed by ESI-TOF MS/MS. Results: The amount of vitreous protein was over 1.5-fold elevated in PDR subjects compared with normal subjects. But protein profile pattern of PDR vitreous body was more simple than control. This result imply fact that protease activity is increased in vitreous. In the normal vitreous, the major protein content consisted of albumin, transferrin, immunoglobulins, α-1-antitrypsin, PEDF, Apo J, prealbumin, as confirmed by a peptide mass fingerprint. PEDF is well known to regulate angiogenesis in eye. Compared to control vitreous, PDR vitreous have lower levels of PEDF, Apo J and transthyretin, but the higher levels of albumin, IgG, IgA, α-1-antitrypsin, complement C3, C4, hemopexin, Apolipoprotein, α-1ß-glycoprotein, and antithrombin III. Conclusions: Although this reflects a shift to a serum-like protein profile resulting from the blood-retinal barrier breakdown, the level of proteins related with inflammation process changed largely. Also, we found the existance levels of two anti-angiogenic factors, PEDF and antithrombin III, changed reciprocally to each other. The present study showed for the first time that antithrombin III exist in the vitreous body of patients with diabetic retinopathy. Moreover, this can explain that blood does not clot within PDR vitreous.
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