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고포도당과 덱사메타존이 섬유주세포의 투과도에 미치는 영향

Effects of High Glucose and Dexamethasone on the Permeability in Trabecular Meshwork Cells

김재우|강선희
대한안과학회 (The Korean Ophthalmological Society)|2018.03|대한안과학회지 (Journal of The Korean Ophthalmological Society)|제59권 제3호|pp. 252-260

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PermeabilityTight junctionDexamethasoneHigh glucoseTrabecular meshwork
주제분류안과학

초록

목적: 인체의 섬유주세포에서 고농도포도당이 섬유주의 투과도와 세포생존에 미치는 영향을 알아보고 이에 대한 덱사메타존의 역할을 알아보고자 한다. 또한 투과도의 변화에 영향을 미치는 섬유주세포의 밀착연접의 변화를 알아보고자 한다. 대상과 방법: 일차배양한 인체의 섬유주세포에 저농도(5 mM)와 고농도(25 mM)의 포도당에 각각 3일간 노출시켰으며 이때 1.0 μM의 덱사메타존에도 함께 노출시켰다. 섬유주단층세포층의 투과도를 carboxyfluorescein permeability test와 transendothelial electrical resistance (TEER)로 각각 측정하였다. 섬유주세포의 접합부에 미치는 영향을 알아보기 위하여 밀착연접의 claudin-5와 occludin의 mRNA와 단백질의 발현을 reverse transcription polymerase chain reaction (RT-PCR)과 western blot으로 각각 측정하였다. 결과: Carboxyfluorescein permeability test와 TEER 결과 모두 고농도포도당은 저농도포도당에 비해 섬유주단층세포층 투과도가 유의하게 크게 나타났으며(p=0.022, 0.028), claudin-5와 occludin mRNA의 발현이 각각 7.5%와 12.9% 작게 나타났다. 덱사메타존에 함께 노출시킨 경우 고농도포도당에 노출시킨 경우에 비해 투과도가 감소되었으며, claudin-5와 occludin mRNA의 발현은 유의한 차이를 나타내지 않았고, claudin-5와 occludin 단백질의 발현은 각각 6.51%, 8.55%로 유의하게 증가하였다(p=0.015, 0.012). 결론: 덱사메타존은 고농도포도당에 의한 섬유주단층세포층의 투과도 증가를 상쇄하였다. 덱사메타존은 섬유주세포의 밀착연접부에 있는 claudin-5와 occludin의 발현을 증가시켰는데 이러한 밀착연접부의 변화는 스테로이드에 의한 섬유주 저항 증가의 기전이 될 수 있을 것으로 생각된다.

Abstract

Purpose: To investigate the effects of high glucose (HG) and dexamethasone (DEX) on the survival and permeability of trabecular meshwork cells (HTMC), and associated changes in tight junctions. Methods: Primary cultured HTMC were exposed to 5 mM low glucose (LG) or 25 mM HG with or without 1.0 μM DEX for 3 days. The permeability of the HTMC monolayer was assessed using carboxyfluorescein or transendothelial electrical resistance (TEER). Gene and protein expressions of claudin-5 and occludin were assessed with reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. Results: HG was significantly associated with greater HTMC monolayer permeability compared to LG by both the carboxyfluorescein permeability test and TEER (p = 0.022, 0.028). HG also decreased claudin-5 and occludin mRNA expression, respectively (7.5%, 12.9%). DEX abolished HG-induced increased permeability, and increased the protein expression of claudin-5 and occludin, respectively (p = 0.015, 0.012). Conclusions: In HTMCs, DEX reversed HG-induced permeability increase. DEX increased tight junction molecules claudin-5 and occludin. Thus, DEX-induced changes in junctional proteins could be another mechanism of increased resistance through the trabecular meshwork and may result in steroid-induced glaucoma.

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