title: Critical role of TNF-α-induced macrophage VEGF and iNOS production in the experimental corneal neovascularization.
abstract: OBJECTIVE
  We evaluated the roles of tumor necrosis factor (TNF)-α in alkali-induced corneal neovascularization (CNV).
  
  
  METHODS
  CNV was induced by alkali injury and compared in wild-type (WT) BALB/c mice, and TNF receptor 1-deficient (TNF-Rp55 KO) counterparts, or in mice treated with TNF-α antagonist and recombinant TNF-α. Angiogenic factor expression and leukocyte accumulation in the early phase after injury were quantified by real-time PCR and immunohistochemical analysis, respectively.
  
  
  RESULTS
  Alkali injury augmented the intraocular mRNA expression of TNF-α and its receptor, together with a transient macrophage and neutrophil infiltration. Compared to WT mice, TNF-Rp55 KO mice exhibited reduced CNV. Intraocular F4/80-positive macrophages and Ly-6G-positive neutrophils infiltration did not change in KO mice compared to WT mice after the injury. Alkali injury induced a massively increased intraocular mRNA expression of angiogenic factors, including vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), interleukin (IL)-6, E-selectin, and intercellular adhesion molecule (ICAM)-1 in WT mice, whereas these increments were retarded severely in KO mice. Immunofluorescence analysis demonstrated that F4/80-positive cells expressed VEGF and iNOS. Moreover, TNF-α enhanced VEGF and iNOS expression by peritoneal macrophage from WT, but not KO mice. Topical application of TNF-α antagonist reduced CNV, while topical application of recombinant TNF-α enhanced it.
  
  
  CONCLUSIONS
  TNF-Rp55-KO mice exhibited impaired alkali-induced CNV through reduced intracorneal infiltrating macrophage VEGF and iNOS expression.
authors.affiliation: Department of Ophthalmology, the First Affiliated Hospital of Soochow University, Suzhou City, China. lupeirong@yahoo.com.hk
authors.forename: Peirong
authors.initials: P
authors.lastname: Lu
authors.forename: Longbiao
authors.initials: L
authors.lastname: Li
authors.forename: Gaoqin
authors.initials: G
authors.lastname: Liu
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Baba
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ishida
authors.forename: Mizuho
authors.initials: M
authors.lastname: Nosaka
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Kondo
authors.forename: Xueguang
authors.initials: X
authors.lastname: Zhang
authors.forename: Naofumi
authors.initials: N
authors.lastname: Mukaida
mesh_terms: D000468:Alkalies / Q000633:toxicity; D000818:Animals; D002478:Cells, Cultured; D003315:Cornea / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D008264:Macrophages / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D000468:Alkalies; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D052247:Nitric Oxide Synthase Type II