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Stimulation of TLR 4 Increases Angiogenic and Anti‐Angiogenic Gene Expression in Choroidal Endothelial Cells
Author(s) -
Stewart E.,
Wei R.,
Uribe M.,
May S.,
Amoaku W.
Publication year - 2015
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2015.0627
Subject(s) - tlr4 , angiogenesis , cxcl10 , inflammation , tlr2 , biology , choroidal neovascularization , chemokine , immunology , stimulation , microarray analysis techniques , microarray , cancer research , medicine , gene expression , retinal , gene , endocrinology , biochemistry
Purpose The Toll‐like receptor ( TLR ) family initiate the innate immune response in reaction to invading microbes. Studies confirm the expression of TLR s in a variety of ocular tissues and cells, and it has also been suggested that selected TLR s may be associated with geographic atrophy and neovascularisation in age‐related macular degeneration, diabetic retinopathy and other vascular and inflammatory diseases of the ocular posterior segment. However, the exact mechanisms of TLR stimulation and the mechanism of endothelial inflammatory and angiogenic effects have not been defined. Methods Microarray analysis was performed on primary human choroidal endothelial cells stimulated with LPS , a specific TLR ligand. Microarray results were validated using qPCR and the functional significance of increased factors were investigated using proliferation and migration assays. Results Microarray indicated a large increase in expression of angiogenic (e.g. CCL 2, IL ‐8) and anti‐angiogenic (e.g. CXCL 10, CXCL 11) molecules. The fold change in expression of these molecules was validated using qPCR and the secretion using ELISA . Reported angiogenic molecules were confirmed to have a pro‐angiogenic effect in vitro. Conclusions This profiling of factors increased during TLR 4 stimulation will help inform our understanding of inflammation in choroidal vascular diseases, as well as the development of future treatments for intraocular vascular diseases.