라타노프로스텐 부노드가 섬유주세포의 투과도에 미치는 영향
Effect of Latanoprostene Bunod on the Permeability of Trabecular Meshwork Cells
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목적: 섬유주세포에서 라타노프로스텐 부노드(latanoprostene bunod, LBN)가 일산화질소(nitric oxide, NO)의 생성과 섬유주의 투과도에 미치는 영향을 알아보고자 하였다. 대상과 방법: 섬유주세포에 50, 100 μM LBN과 라타노프로스트 유리산(latanoprost free acid, LAT)을 30분간 노출시켰으며, 0.5 mM L-NAME (N-Nitroarginine methyl ester)에도 함께 노출시켰다. 세포 생존과 NO 생성은 MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay와 Griess assay로 측정하였다. 섬유주단층세포층의 투과도와 저항도는 carboxyfluorescein permeability와 trans-endothelial electrical resistance (TEER)로 각각 측정하였다. 결과: 100 μM LBN은 NO의 생성을 증가시켰으며, L-NAME에 함께 노출시킨 경우에는 NO 생성이 억제되었다. 100 μM LBN은 섬유주단층세포층의 TEER을 저하시켰다. 또한 carboxyfluorescein permeability test에서 LBN은 투과도를 증가시켰으며, 100 μM LBN에L-NAME를 함께 노출시킨 경우 투과도가 감소하였다. LAT는 NO의 생성과 섬유주단층세포층의 투과도에 영향을 미치지 않았다. 결론: 섬유주세포에서 LBN은 NO의 생성을 증가시키며, 섬유주단층세포층의 투과도를 촉진시켰다. 따라서 LBN은 포도막공막유출의증가와 함께 섬유주를 통한 방수유출을 증가시키는 작용을 나타낼 것으로 생각된다.
Abstract
Purpose: To investigate the effects of latanoprostene bunod (LBN) on nitric oxide (NO) production and permeability in human trabecular meshwork cells (HTMC). Methods: HTMC were treated with 50 and 100 μM LBN and latanoprost free acid (LAT) for 30 minutes. Additionally, 100 μM LBN was co-exposed to 0.5 mM L-NAME (N-Nitroarginine methyl ester). Cellular viability and NO production were measured using MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) and Griess assays. The permeability and resistance of the HTMC monolayer were evaluated by trans-endothelial electrical resistance (TEER) and carboxyfluorescein permeability. Results: Exposure to 100 μM LBN led to increased NO production, whereas co-exposure to L-NAME reduced NO production. Treatment with 100 μM LBN decreased the TEER of the HTMC monolayer. LBN exposure heightened carboxyfluorescein permeability, but co-exposure to 100 μM LBN and L-NAME reduced permeability. LAT treatment did not affect NO production or permeability. Conclusions: LBN increased the permeability of the HTMC monolayer and increased NO production. Therefore, LBN might increase trabecular outflow in addition to promoting uveoscleral outflow.
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