Abstract
Purpose: :
To investigate whether oxidative stress-induced changes in human trabecular meshwork (TM) cells, as seen in primary open angle glaucoma, can be prevented by treatment with omega-3 or omega-6 fatty acids.
Methods: :
TM cells were isolated from 5 donors. Cultures of passages 3 to 5 were treated with 50µM omega-3 or 5pg/ml omega-6 fatty acids in serum-free Ham`s F10 for 48 hours. Subsequently, oxidative stress was induced by exposure to 300µM hydrogen peroxide (H2O2) for 1 hour. Influence of omega-3 and omega-6 fatty acids on expression levels of fibronectin-1 (FN-1), heat-shock protein-27 (HSP-27) and plasminogen activator inhibitor-1 (PAI-1) mRNA were analyzed and quantified by real-time PCR. To examine the expression of extracellular matrix (ECM) proteins and oxidative stress-induced proteins, TM cells and their medium were separately assayed with a RayBio® Human Cytokine Antibody Array C Series 4000. Immunofluorescence labeling was used to analyze the influence of omega-3 and omega-6 fatty acids on expression of connective tissue growth factor (CTGF), FN-1, PAI-1, HSP-27 and HSP-90.
Results: :
Treatment with omega-3 and omega-6 fatty acids decreased the levels of FN-1, PAI-1 and HSP-27 mRNA in real-time PCR. The repressive effect of omega-6 fatty acids on the analyzed oxidative stress-induced proteins was stronger and more pleiotropic than that of omega-3 fatty acids. This was confirmed by cytokine antibody array analysis of TM cells and their corresponding medium. By immunofluorescence analysis TM cells treated with omega-3 or omega-6 fatty acids displayed decreased labeling intensities for CTGF, FN-1, PAI-1, HSP-27 and HSP-90.
Conclusions: :
Omega-3 and omega-6 fatty acids both had antioxidative effects on oxidative stress exposed human TM cells. Omega-6 fatty acids had a stronger repressive antioxidative effect, and targets of Omega-6 and Omega-3 fatty acids were different to some extent. Therefore, a combinatory treatment might maximize the protective effect.
Keywords: trabecular meshwork • oxidation/oxidative or free radical damage