Epigallocatechin gallate enhances human lens epithelial cell survival after UVB irradiation via the mitochondrial signaling pathway.

Abstract

The aim of the present study was to explore the mechanism underlying the ultraviolet B (UVB) irradiation-induced apoptosis of human lens epithelial cells (HLECs), and to investigate the protective effect of epigallocatechin gallate (EGCG) against the UVB-induced apoptosis of HLECs. HLECs were exposed to different concentrations of EGCG plus UVB (30 mJ/cm2). Cell viability was determined using the MTT assay. Furthermore, mitochondrial membrane potential (Δψm) and apoptosis were assessed by flow cytometry with JC-1 and Annexin V/PI staining, respectively. Moreover, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), as well as the levels of GSH, hydrogen peroxide (H2O2) and hydroxyl free radicals were determined using biochemical assay techniques. Reverse transcription-quantitative PCR and western blotting were used to detect the mRNA and protein expression levels of Bcl-2, Bax, cytochrome c, caspase-9 and caspase-3, respectively. The results revealed that UVB irradiation reduced the Δψm of HLECs and induced apoptosis. Notably, EGCG significantly attenuated the generation of H2O2 and hydroxyl free radicals caused by UVB irradiation in HLECs, and significantly increased CAT, SOD and GSH-Px activities, however, the GSH levels were not significantly increased. EGCG also reduced UVB-stimulated Bax, cytochrome c, caspase-9 and caspase-3 expression, and elevated Bcl-2 expression, suggesting that EGCG may possess free radical-scavenging properties, thus increasing cell viability. In conclusion, EGCG may be able to protect against UVB-induced HLECs apoptosis through the mitochondria-mediated apoptotic signaling pathway, indicating its potential application in clinical practice.