Protection of Mcc950 against H2O2-induced inflammatory injury in ARPE-19 cells / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

@#AIM: To research the protection of Mcc950, the inhibitor of NLRP3, against the inflammatory injury to human retinal pigment epithelium cell line ARPE-19. <p>METHODS: Cultured cell line ARPE-19 was divided into control group, H₂O₂ treating group, Mcc950 treating group and Mcc950+H₂O₂ treating group. Different concentrations of H₂O₂ and Mcc950 were used to treat the cells. Cell activity was detected by using CKK8 and proper experimental concentration of H₂O₂ and Mcc950 were determined. After the treatment, the concentration of IL-1β were detected by using ELISA. The change of NLRP3 related proteins were detected by Western blot. And cell apoptosis was examined by TUNEL stain. <p>RESULTS: Cell ability was gradually decreased along with the increasing treating concentrations of H₂O₂. Cell ability showed statistical difference when the concentration of H₂O₂ arrived 400μmol/L. With the concentration of 0.1 and 1μmol/L, Mcc950 had no effect on cell ability. So we chose 400μmol/L H₂O₂ and 1μmol/L Mcc950 as the experimental concentrations. Compared with the normal control group, the cell viability in the H₂O₂ treating group was significantly reduced, the IL-1β in the supernatant was significantly increased, and the apoptosis rate was significantly increased, with statistically significant differences(<i>P</i><0.05). In Mcc950+H₂O₂ treating group, cell viability was significantly increased, the IL-1β in the supernatant and the apoptosis rate were significantly decreased(<i>P</i><0.05). By Western blot, after treated with 400μmol/L H₂O₂, the IL-1β, NLRP3, pro-caspase1 and caspase1 were obviously increased compared to control group. After treated with Mcc950, the NLRP3 and pro-caspase1 still were at high level, the expression of caspase1 was suppressed, which indicated that Mcc950 effectively inhibited the activation of NLRP3 inflammasome consequently disturbed the formation of caspase1.<p>CONCLUSION: Mcc950 can inhibits the function of NLRP3, leading to increasing of the cell ability and decreasing of the cell apoptosis.