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Naoko Tsuboi, Toshihiro Inoue, Motofumi Kawai, Miyuki Inoue-Mochita, Tomokazu Fujimoto, Nanako Awai-Kasaoka, Akitoshi Yoshida, Hidenobu Tanihara; The Effect of Monocyte Chemoattractant Protein-1/CC Chemokine Ligand 2 on Aqueous Humor Outflow Facility. Invest. Ophthalmol. Vis. Sci. 2012;53(10):6702-6707. doi: 10.1167/iovs.12-10376.
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To investigate the effect of monocyte chemoattractant protein-1 (MCP-1)/CC chemokine ligand 2 on aqueous humor outflow facility.
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.
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.
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.
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