Sirpa Loukovaara1, Peppi Koivunen, Marta Inglés, Javier Escobar, Maximo Vento, Sture Andersson. 1. Unit of Vitreoretinal Surgery, Department of Ophthalmology, Helsinki University Central Hospital, Helsinki, FinlandOulu Center for Cell-Matrix Research, Biocenter Oulu, Department of Medical Biochemistry and Molecular Biology, Oulu University, Oulu, FinlandDepartment of Physiology, University of Valencia, Spain, Valencia, SpainNeonatal Research Unit & Division of Neonatology, University and Polytechnic Hospital & Health Research Institute La Fe, Valencia, SpainDepartment of Pediatrics, Helsinki University Central Hospital, Helsinki, Finland.
Abstract
PURPOSE: To evaluate the role of protein carbonyls and hypoxia inducible factor-1α (HIF-1α) in diabetic eyes with proliferative diabetic retinopathy (PDR). METHODS: Prospective consecutive controlled observational study was performed. Vitreous samples were collected at the start of the 3-ppp vitrectomy. Protein carbonylation analysis was performed by Western blotting with antibody against 2,4-Dinitrophenol (anti-DNP), following derivatization of protein carbonyls with 2,4 Dinitrophenylhydrazine (DNHP). Protein carbonylation was quantified by scanning densitometry analysis and relativized to the total amount of protein into the ponceau staining of membranes. Vitreous HIF-1 α was determined with ELISA in a subgroup of the samples. Thirty-one eyes were operated due to PDR (study group). Of the 189 controls, 39 had nonproliferative diabetic retinopathy (non-PDR), 111 retinal detachment (RD) and 39 macular hole/pucker (MH). RESULTS: Comparison of eyes with PDR with controls revealed that the mean vitreous concentrations of protein carbonyls were significantly higher in the eyes affected with PDR being 242±130 (SD) compared with non-PDR controls 180±142, nondiabetic eyes affected with RD 175±131 and MH/pucker 140±95 (p=0.008, one-way anova). Mean HIF-1α values were higher in eyes with PDR compared with controls (RD, MH/pucker); the values being 0.53±0.34 (SEM; n=4) and 0.13±0.04 (SEM; n=19), respectively (p=0.009). CONCLUSIONS: Protein carbonyl and HIF-1 α levels were significantly increased in the vitreous fluid of surgically treated eyes with PDR. Our findings suggest an association between increased intravitreal levels of protein carbonyls and the pathogenesis of PDR.
PURPOSE: To evaluate the role of protein carbonyls and hypoxia inducible factor-1α (HIF-1α) in diabetic eyes with proliferative diabetic retinopathy (PDR). METHODS: Prospective consecutive controlled observational study was performed. Vitreous samples were collected at the start of the 3-ppp vitrectomy. Protein carbonylation analysis was performed by Western blotting with antibody against 2,4-Dinitrophenol (anti-DNP), following derivatization of protein carbonyls with 2,4 Dinitrophenylhydrazine (DNHP). Protein carbonylation was quantified by scanning densitometry analysis and relativized to the total amount of protein into the ponceau staining of membranes. Vitreous HIF-1 α was determined with ELISA in a subgroup of the samples. Thirty-one eyes were operated due to PDR (study group). Of the 189 controls, 39 had nonproliferative diabetic retinopathy (non-PDR), 111 retinal detachment (RD) and 39 macular hole/pucker (MH). RESULTS: Comparison of eyes with PDR with controls revealed that the mean vitreous concentrations of protein carbonyls were significantly higher in the eyes affected with PDR being 242±130 (SD) compared with non-PDR controls 180±142, nondiabetic eyes affected with RD 175±131 and MH/pucker 140±95 (p=0.008, one-way anova). Mean HIF-1α values were higher in eyes with PDR compared with controls (RD, MH/pucker); the values being 0.53±0.34 (SEM; n=4) and 0.13±0.04 (SEM; n=19), respectively (p=0.009). CONCLUSIONS: Protein carbonyl and HIF-1 α levels were significantly increased in the vitreous fluid of surgically treated eyes with PDR. Our findings suggest an association between increased intravitreal levels of protein carbonyls and the pathogenesis of PDR.
Authors: Ghulam Mohammad; Kaiser Alam; Mohammad Imtiaz Nawaz; Mohammad Mairaj Siddiquei; Ahmed Mousa; Ahmed M Abu El-Asrar Journal: J Physiol Biochem Date: 2015-06-04 Impact factor: 4.158