Puerarin alleviates hyperosmotic stress-induced oxidative stress, inflammation, apoptosis and barrier damage of human corneal epithelial cells by targeting SIRT1/NLRP3 signaling

The increase of tear osmolarity caused by excessive evaporation of tear phase is the main pathological mechanism of dry eye disease (DED). Puerarin, the major bioactive ingredient isolated from the root of the Pueraria lobata (Willd.) Ohwi, has been reported to improve ophthalmic diseases in clinic. However, the effect and the potential regulatory mechanism related to silent information regulator sirtuin 1 (SIRT1)/NOD-like receptor family pyrin domain containing 3 (NLRP3) signaling of puerarin in DED has not been evaluated. In this study, we aimed to explore the effect and mechanism of hyperosmotic stress (Hyp)-induced human corneal epithelial cell line (HCE-2). The viability of HCE-2 cells induced by Hyp with or without puerarin treatment was assessed by a CCK-8 assay. Results indicated that puerarin treatment enhanced cell viability, reduced reactive oxygen species (ROS) content, increased CAT and SOD activities, and elevated the ratio of GSH/GSSG in HCE-2 cells exposed to Hyp. Besides, TNF-α, IL-1β and IL-6 contents were decreased by puerarin. Additionally, puerarin inhibited Hyp-induced apoptosis and barrier disruption of HCE-2 cells. Moreover, molecular docking method suggested that puerarin bound to SIRT1, and upregulated SIRT1 and downregulated NLRP3 inflammasome proteins after puerarin treatment was observed. Furthermore, SIRT1 silencing alleviated the protective effects of puerarin on Hyp-induced HCE-2 cell damage. Collectively, puerarin attenuates Hyp-induced injury of HCE-2 cells by targeting regulating SIRT1/NLRP3 signaling.