April 2014
Volume 55, Issue 13
ARVO Annual Meeting Abstract  |   April 2014
Minocycline abrogates glycated albumin induced MCP-1 and IL-8 cytokine release from ARPE-19 cells
Author Affiliations & Notes
  • Joanna DaCosta
    Cranfield University, Bedfordshire, United Kingdom
  • Footnotes
    Commercial Relationships Joanna DaCosta, None
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Investigative Ophthalmology & Visual Science April 2014, Vol.55, 4953. doi:
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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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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: 412 age-related macular degeneration • 490 cytokines/chemokines • 557 inflammation  

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