수정체상피세포에서 Ceramide 및 Phytoceramide에 의해 유발된 세포자멸사

Abstract

Purpose: The purpose of this study was to determine if ceramide, which is known as secondary messenger of programmed cell death (apoptosis), can cause apoptosis in lens epithelial cell (LEC) and if so, to identify the pathway by which apoptosis occurs. Methods: After LECs were exposed to various concentrations of ceramide and phytoceramide, we evaluated the resulting apoptosis response using the Hoechst-EthD stain and Annexin stain. To search for the apoptosis pathway, LECs were preincubated in various concentrations of CPP32-like protease inhibitor, specific caspase-8 inhibitor, and specific caspase-9 inhibitor, then treated with ceramide and phytoceramide. We performed LDH assay 12 hours later. Cytochrome c immunostaining was done after exposure to the ceramide and phytoceramide. Results: All kinds of ceramide induced time and concentration dependent apoptosis m LEC. Caspase 8 inhibitor and caspase 9 inhibitor reduced the apoptosis in ceramide VI, phytoceramide II, and phytoceramide VI. In all ceramides, cytochrome c staining was positive. Conclusions: Ceramide and phytoceramide can cause apoptosis in LEC. Ceramide and phytoceramide may be used to prevent the posterior capsular opacity after cataract surgery.