Figure 7

Fisetin inhibits the formation of NLRP3 inflammasome and the expression of IL-1β in zebrafish larvae. After 3 days of fertilization, zebrafish larvae (n = 20) were pretreated with the indicated concentrations of fisetin for 2 h prior to stimulation with 5 µg/mL LPS for 2 h, and subsequent stimulation with 2 mM ATP (LPS/ATP). (A) After 24 h, heart rates were measured for 2 min and expressed as beats/min. (B) Neutral red staining for detecting the distribution of macrophages after 24 h of treatment with LPS/ATP. (C) Total RNA was extracted after 18 h of treatment with LPS/ATP, and the expression of IL-1β, ASC, caspase-A, and caspase-B was determined using RT-PCR. β-Actin was used as a loading control. Relative densities were calculated using ImageJ software. The results indicate the mean ± standard error median (SEM), and is representative of the results obtained from three independent experiments.^# p < 0.05 vs. untreated zebrafish larvae; *** p < 0.001 and * p < 0.05 vs. LPS/ATP-treated zebrafish larvae.