Effects of Low-Dose Cyclosporine on Human Corneal Epithelial Cells. |
Sang Jun Lee, Su Jin Kim, Jong Soo Lee |
1Department of Ophthalmology, Pusan National University School of Medicine, Busan, Korea. jongsool@pusan.ac.kr 2Department of of Ophthalmology, Maryknoll Medical Center, Busan, Korea. |
저농도에 따른 사이클로스포린의 인체 각막상피세포에 미치는 영향 |
이상준1⋅김수진2⋅이종수1 |
Department of Ophthalmology, Pusan National University School of Medicine1, Busan, Korea Department of of Ophthalmology, Maryknoll Medical Center2, Busan, Korea |
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Abstract |
PURPOSE To evaluate the response and cellular changes of cultured human corneal epithelium according to the concentration of topical cyclosporine. METHODS: Human corneal epithelial cells were exposed to cyclosporine A concentrations of 1 ug/ml (0.0001%), 10 ug/ml (0.001%), 100 ug/ml (0.01%), and 500 ug/ml (0.05%) for 5 and 10 minutes. An MTT-based colorimetric assay was performed to assess the metabolic activity of cellular proliferation and a lactate dehydrogenase (LDH) leakage assay was used to determine cellular toxicity. The modulations of extracellular matrix proteins such as PIP and laminin were evaluated. The levels of pro-inflammatory cytokine, TNF-alpha, and IL-1 were evaluated by ELISA kits. RESULTS: The inhibitory effects of human corneal epithelial cellular proliferation did not show a concentration- or exposure time-dependent response. Activity of LDH did not show a statistically significant difference for the different concentrations and exposure times. The inhibitory effects of extracellular matrix proteins such as PIP or laminin synthesized from human corneal epithelial cells were compared with those in the control group and showed a statistically significant difference at a cyclosporine concentration greater than 0.01%. Released TNF-alpha and IL-1 from human corneal epithelial cells did not show any difference according to concentration or cyclosporine exposure time. CONCLUSIONS: To modulate human corneal epithelial cellular proliferation and the levels of extracellular matrix proteins such as PIP and laminin, a concentration of cyclosporine A greater than 0.01% for longer than 5 minutes of exposure is needed. There were little effects of cyclosporine A on pro-inflammatory cytokine secreted from corneal epithelial cells. |
Key Words:
Corneal epithelium;Cyclosporine;Toxicity |
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