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104. Jahrestagung der Deutschen Ophthalmologischen Gesellschaft 2006
Abstract
Abstract
SA.11.09 Effects of sera from glaucoma patients on the survival and protein expression in retinal ganglion cells (RGC) Bell K.1, Seigel G.2, Pfeiffer N.1, Grus F. H.1
1Department of Ophthalmology, University of Mainz, Germany; 2Ross Eye Institute, University of Buffalo, USA Objective: Recent studies could show complex antibody profiles against retinal and optic nerve antigens in glaucoma patients. It was the aim of our study to analyze if sera of patients with glaucoma show effects on the death and protein expressions of RGC.
Methods: The retinal ganglion cells line R28 were exposed to DMEM with either 10% serum from patients with glaucoma or from healthy subjects. The cells were exposed to an elevated pressure of 15000 Pa (112mmHg) for 48 h. Control cells were incubated at atmospheric pressure. After lysis, the protein expression profiles were measured by SELDI-TOF, Ciphergen, USA using two different protein chips (CM10/H50). These profiles were analyzed by multivariate statistics to find the most significant biomarkers. Cell viability was rated with cell proliferation reagent WST-1(Roche).
Results: In this pilot experiment, the cells with glaucoma sera and pressure showed a decrease in cell viability compared to those cells incubated with healthy sera. Additionally we could show modified protein expression profiles. Around 400 different protein and peptide cluster could be detected by Seldi-TOF. From these, 6 protein biomarkers were calculated by the analysis of discriminance which were expressed significantly different between the treatment groups (WilksÂ’ l=0.0289 P<0.001). The Mahalanobis distances (Md) were calculated which revealed the highest distance from controls in the cells which were incubated with glaucoma sera at elevated pressure (Md=40; P<0.0001). A Variance Component and Mixed model ANOVA was calculated to assess the effect of pressure and type of sera on the protein expression profiles. This analysis showed no significant effect of high pressure or sera only, but a highly significant effect (P<0.001) of the combined effect of elevated pressure by type of sera.
Conclusions: The cells incubated with serum from glaucomatous patients showed a significantly different protein expression in comparison to those cells that were incubated with healthy serum. This effect was clearer if the cells were incubated under elevated pressure. Considering that antiretinal antibodies are present in the glaucoma sera, this pilot study might help to understand the complex interactions of sera and elevated pressure on the apoptosis of RGC and might provide hints for the increased liability of RGC to elevated intraocular pressure.
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