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Cholestanol Induces Apoptosis of Corneal Endothelial and Lens Epithelial Cells

¹ From the Department of Physiological Chemistry and Metabolism, Graduate School of Medicine, and ² Department of Ophthalmology, Branch Hospital, The University of Tokyo, Japan; and the ³ Department of Ophthalmology, Jichi Medical School, Tochigi, Japan.

PURPOSE. To determine whether cholestanol induces cornea endothelial and lens epithelial cell death in vitro.

METHODS. Cornea endothelial and lens epithelial cells were cultured in minimum essential media with 10% fetal bovine serum containing 10 µg/ml cholesterol in ethanol, 10 µg/ml cholestanol in ethanol, or 1% ethanol. These cells, stained using the terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) method, were analyzed by laser cytometer. The activities of ICE and CPP32 proteases in cells were also measured.

RESULTS. Both cornea endothelial and lens epithelial cells cultured with 10 µg/ml cholestanol showed a significant loss of viability. The nuclei of these cells cultured with 10 µg/ml cholestanol were more frequently stained than those exposed to 10 µg/ml cholesterol or 1% ethanol. Quantitative analysis of apoptotic DNA fragmentation confirmed that the cholestanol induced apoptosis of these cells in a time-dependent manner. The activities of interleukin-1ß–converting enzyme (ICE) and CPP32 proteases for cells cultured with 10 µg/ml cholestanol were significantly higher than those observed in control cells.

CONCLUSIONS. In vitro, cholestanol was taken up by corneal endothelial cells and lens epithelial cells, an event that led to apoptosis of these cells.

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