title: The effect of monocyte chemoattractant protein-1/CC chemokine ligand 2 on aqueous humor outflow facility.
abstract: OBJECTIVE
  To investigate the effect of monocyte chemoattractant protein-1 (MCP-1)/CC chemokine ligand 2 on aqueous humor outflow facility.
  
  
  METHODS
  Aqueous humor outflow facility was measured in enucleated porcine eyes in a constant pressure perfusion system with or without MCP-1 (1600 ng/mL). Expression of CCR2, an MCP-1 receptor, in Schlemm's canal endothelial (SCE) cells was examined by reverse transcription-polymerase chain reaction (RT-PCR) assay. The effect of MCP-1 (0-1600 ng/mL) on SCE cell viability was evaluated using a WST-8 assay. The effect of MCP-1 (0-800 ng/mL) on SCE-cell monolayer permeability was evaluated with or without a CCR2 antagonist (10 nM) by measuring transendothelial electrical resistance (TEER). The intracellular localization of the gap junction protein ZO-1 was analyzed by immunofluorescence staining of SCE cells.
  
  
  RESULTS
  The aqueous humor outflow facility increased significantly from basal levels at 80 minutes after perfusion with MCP-1 compared with control eyes (21.2% ± 6.6% [MCP-1] vs. 5.7 ± 2.5% [control]; P = 0.048). CCR2 was detected by RT-PCR. Cell viability was not affected by MCP-1 treatment. TEER of SCE-cell monolayer at 3 hours after treatment with 800 ng/mL MCP-1 decreased by 21.6 ± 1.7% compared with controls (P = 0.014), and the TEER-decreasing effects of MCP-1 were attenuated by a CCR2 antagonist. Immunocytochemical staining revealed a modest disruption of ZO-1 in MCP-1-treated SCE cells.
  
  
  CONCLUSIONS
  The present results revealed that MCP-1 increased aqueous humor outflow facility and decreased TEER via CCR2. These findings suggest that MCP-1 modulates aqueous humor outflow through the conventional pathway.
authors.affiliation: Department of Ophthalmology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Tsuboi
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Inoue
authors.forename: Motofumi
authors.initials: M
authors.lastname: Kawai
authors.forename: Miyuki
authors.initials: M
authors.lastname: Inoue-Mochita
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Fujimoto
authors.forename: Nanako
authors.initials: N
authors.lastname: Awai-Kasaoka
authors.forename: Akitoshi
authors.initials: A
authors.lastname: Yoshida
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Tanihara
mesh_terms: D000818:Animals; D001082:Aqueous Humor / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000494:pharmacology*; D007150:Immunohistochemistry; D013552:Swine; D014129:Trabecular Meshwork / Q000166:cytology / Q000187:drug effects / Q000502:physiology*
chemical_list: D018932:Chemokine CCL2