Abstract:AIM:To investigate the protective effects of resveratrol(RSV)on inflammation and oxidative stress damage in human corneal epithelial cells(HCECs).
METHODS: The inflammation of HCECs was induced by Tumor necrosis factor-α(TNF-α), and the experiment was divided into: control group, TNF-α group and RSV+TNF-α group. The oxidative stress response of HCECs was induced by H2O2, and they were divided into normal group, H2O2 group and RSV+ H2O2 group. MTT assay was used to detect the viability of HCECs; RT-qPCR and enzyme-linked immunosorbent assay(ELISA)methods were used to detect the expression of IL-1, IL-6 and IL-8; Immunofluorescence staining and Western blot were used to observe the nuclear translocation of NF-κB p65. 2',7'-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe was applied to detect the level of reactive oxygen species(ROS).
RESULTS:In the inflammatory response of HCECs, RT-qPCR and ELISA showed that the expression levels of IL-1, IL-6 and IL-8 were increased significantly in the TNF-α group compared with the control group, the above indicators were lower after pretreatment of RSV than those in TNF-α group; Immunofluorescence staining and Western blot showed that the nuclear translocation of NF-κB p65 was increased in TNF-α group, while it was inhibited after pretreatment of RSV. In the oxidative stress response of HCECs, the results of MTT and DCFH-DA fluorescent probe staining showed that H2O2 significantly decreased the viability of HCECs and increased the production of ROS in HCECs. After pretreatment of RSV, cell viability increased significantly, and RSV inhibited the generation of ROS in HCECs induced by H2O2.
CONCLUSION: RSV has an inhibitory effect on inflammation and oxidative stress damage in human corneal epithelial cells, and it has been confirmed that RSV inhibits inflammation by inhibiting the activation of the NF-κB pathway.
