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Joanna DaCosta; Minocycline abrogates glycated albumin induced MCP-1 and IL-8 cytokine release from ARPE-19 cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4953.
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Reduced choroidal blood flow may create hypoxia in the retina, triggering formation of new vessels through the activation of vascular endothelial growth factor. This may initiate neovascular age-related macular degeneration.Glycated human serum albumin (GHSA) has been shown to stimulate RPE cells to produce IL-8 and MCP-1. Early glycated albumin may have a direct impact on cell function during ageing. Minocycline is a semisynthetic derivative of tetracycline with a longer half life and improved penetration through the blood brain barrier. Apart from antimicrobial effects it also has potent anti-inflammatory and immunomodulatory effects. It was postulated that minocyline may suppress the induced expresion of inflammatory cytokines from retinal pigment epithelial cells in culture.
Research was conducted on human ARPE-19 cells in culture. Cells were exposed to hypoxia and GHSA. Minocycline was investigated for the effect on cell growth and cell viability. Cell counts were obtained with a Scepter cell counter (Millipore). Cell viability and apoptosis rates were investigated with annexin V and propidium iodide staining by flow cytometry. The effects of hypoxia and different concentrations of GHSA, IL-1β, TNF-α on cell viability and with treatment with minocycline were investigated. ELISA for IL-8 and MCP-1 was conducted with appropriate negative and positive controls. Standards and samples were analysed in duplicate. Intraplate and interplate reproducibility tests were performed.
Cell viability decreased at minocycline doses above 5µM. Hypoxia increased the proportion of non viable cells and cells undergoing apoptosis and minocycline treated cells had a lower proportion in the early stage of apoptosis. ELISA results demonstrated that hypoxia and GHSA treated cells showed a significantly increased MCP-1 production. Minocycline significantly reduced MCP-1 production in normoxic conditions (p< 0.01)and also decreased MCP-1 production in hypoxia (p<0.05). For IL-8, minocycline significantly reduced production in normoxia (p<0.01) but not statistically significantly under hypoxia.
The results suggest that minocycline abrogates the production of inflammatory cytokines IL-8 and MCP-1 in cell culture and minocycline may have a therapeutic role in the treatment of inflammatory changes in age related macular degeneration.
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