Calcium facilitates NLRP3 inflammasome-induced oxidative stress in SHSY5Y cells

Abstract

Objective To study calcium chelator BAPTA-AM antagonize the cellular oxidative stress induced by hydrogen peroxide (H2O2)and to explore the effect of calcium ion on the cell degeneration mediated by NLRP3. Methods The SHSY5Y cell model of oxidative stress was made by hydrogen peroxide, then the cell model was treated with calcium ion carrier A23187 or BAPTA-AM, a higher efficiency calcium chelating agent. The cells were divided into 4 groups: H2O2 treatment group, H2O2+ A23187 group, H2O2 + A23187 + BAPTA-AM group and control group. NLRP3 protein was detected by Western blot, and Caspase-1 and IL-1β were detected by ELISA. Results NLRP3 expression was significantly increased in cells treated by hydrogen peroxide(P<0.05). The NLRP3 protein continued to increase, and the expression of Caspase-1((57.1±19.2)pmol/L) and IL-1β((484.2±49.5)pg/ml) protein was also increased significantly in cells treated by A23187, and the difference had statistically significant for caspase-1 or IL-1β in H2O2+ A23187 group compared with those in control group(Caspase-1: (26.8±12.9)pmol/L, IL-1β: (326.9±52.1)pg/ml)(P<0.05, P<0.01). NLRP3, Caspase-1 and IL-1β were all significantly reduced after adding a higher efficiency calcium chelating agent BAPTA-AM. Conclusion Calcium overload is likely to enhance the oxidative stress induced by hydrogen peroxide and engender neurodegeneration mediated by NLRP3 inflammasome. Key words: Hydrogen peroxide; Caspase-1; Nucleotide binding to the receptor protein 3; Calcium ion