Abstract

To explore the protective function and molecular mechanism of tribuloside in hydrogen peroxide-induced apoptosis and oxidative damage of human lens epithelial cells. Cells were divided into control group, hydrogen peroxide group, hydrogen peroxide+tribuloside 1 μg/ml group, hydrogen peroxide+tribuloside 3 μg/ml group, hydrogen peroxide+tribuloside 10 μg/ml group, hydrogen peroxide+microRNA-NC group, hydrogen peroxide+microRNA-335-3p group, hydrogen peroxide+tribuloside+anti-microRNA-335-3p group. Cell counting kit-8 method and flow cytometry were applied for determining cell viability and apoptosis. Malondialdehyde level, catalase and superoxide dismutase activities were determined using kits. Reverse transcription-quantitative polymerase chain reaction was used for microRNA-335-3p quantification. Kriippellike factor 6 expression was examined via Western blot. Target regulation relationship of miR-335-3p and Kriippel-like factor 6 was analyzed via dual-luciferase report assay. Contrasted with the control group, cell viability, catalase and superoxide dismutase viabilities, and microRNA-335-3p expression were significantly reduced in hydrogen peroxide group (p<0.05), while apoptosis, malondialdehyde and Kriippel-like factor 6 levels were overtly promoted (p<0.05). Contrasted to hydrogen peroxide group, cell viability, catalase and superoxide dismutase activities, and microRNA-335-3p expression in hydrogen peroxide+tribuloside 1 μg/ml group, hydrogen peroxide+tribuloside 3 μg/ml group, hydrogen peroxide+tribuloside 10 μg/ml group were obviously enhanced (p<0.05), but cell apoptosis, malondialdehyde level and Kriippel-like factor 6 protein level were suppressed (p<0.05). Relative to hydrogen peroxide+microRNA-NC group, cell viability, catalase and superoxide dismutase activities in hydrogen peroxide+microRNA-335-3p group were markedly elevated (p<0.05), whereas apoptosis inhibition, malondialdehyde level reduction and Kriippel-like factor 6 protein down-regulation were induced (p<0.05). Compared with hydrogen peroxide+tribuloside group, cell viability, catalase and superoxide dismutase activities in hydrogen peroxide+tribuloside+anti-microRNA-335-3p group were signally inhibited (p<0.05), but apoptosis, malondialdehyde and Kriippel-like factor 6 levels were accelerated (p<0.05). MicroRNA-335-3p directly interacted with Kriippel-like factor 6. Tribuloside could attenuate apoptosis and oxidative damage in hydrogen peroxide-induced cataract model, which was related to microRNA-335-3p/Kriippel-like factor 6 axis.