Abstract
Purpose :
In this study, we investigated the mechanism underlying the HUA-induced damages on retinal epithelium cells by using the ARPE-19 cells under in vitro conditions.
Methods :
The effects of UA (uric acid) concentrations at 4, 8, and 12 mg/dl on cell proliferation as detected by CCK-8 assay were detected at 0, 24, 48, and 72 hours. The impact of high glucose and high osmolality along with high UA was also investigated. Western blot analysis and immunofluorescence microscopy were performed for NLRP3 and caspase 1, along with Annexin A1 expression. The intra- and extracellular milleu were characterized with high oxidative stress as indicated by trolox equivalent antioxidant capacity assay (TEAC) and thiobarbituric acid reactive substances (TBARS) assays.
Results :
Significantly reduced cell proliferation was detected, starting at 24 hours for 12mg/dl UA and at 48 hours for 8 and 12mg/dl UA. However, the damages caused by HUA did not reverse when the UA concentration was returned to 0 mg/dl at 24 hours after exposure. Besides, high glucose and high osmolality could aggravate the damages caused by HUA. Western blot analysis and immunofluorescence microscopy revealed elevation of NLRP3 and caspase 1, along with Annexin A1 dysregulation. The extracellular milleu were characterized with high oxidative stress, contrary to the intra-cellular status.
Conclusions :
In conclusion, HUA could damage ARPE-19 cells through NLRP3 inflammasome activation caused by dual intra- and extra-cellular events that converge together.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.