Human lens epithelial cells ; Nuclear factor kappa B ; Ultraviolet irradiation
Abstract
Purpose: This study was performed to determine the role of nuclear factor kappa B (NF-κ B) on the lens epithelial cell death after ultraviolet (UV) irradiation.
Methods: Simian virus 40 transfected human lens epithelial cells (HLE B-3 cells) were used in this study. UVB located at 10cm from the bottom was irradiated during 1, 2, 3 and 4 minutes. To measure the cytotoxicity N= .assay was used. Translocation of NF-x B was examined by immunocytochemistry with anti NF-κ B p65 antibody and electrophoretic mobility shift assay (EMSA). To confirm the role of NF-κ B, the cells were pretreated with sulfasalazine, a specific inhibitor of NF-κ B, for 30 minutes before irradiation, and cytotoxicity and translocation of NF-κ B were evaluated. .
Results: UV irradiation produced a progressive cytotoxic effect in cultured HLE B-3 cells after 1 minute and maximum cytotoxicity was reached after 3 minutes irradiation. When HLE B-3 cells were irradia´ted with UVB, the translocation of NF-κ B was observed in immunocytochemistry. These translocations were peaked 6 hours after UV irradiation in EMSA. In HLE B-3 cells pretreated with sulfasalazine, the translocation of NF-κ B was blocked. The cellular death after UV irradiation was markedly blocked by sulfasalazine. UV irradiation can translocate NF-κ B and sulfasalazine is a useful blocking agent in this pathway. In addition, sulfasalazine can prevent cellular death after UV irradiation.
Conclusions: These findings suggest that NF-κ B plays an important role in cellular death after UV irradiation.