To confirm the above results, we measured the levels of IL-1β and IL-18 in the medium of astrocytes incubated for 24 h with ATP or LPS or ethanol (10 and 50 mM), in the presence or absence of Mito-TEMPO or z-YVAD-FMK or Z-VAD-FMK or NLRP3 bp. According to the above results, ATP or LPS or ethanol (10 and 50 mM) treatment induces the production of IL-1β and IL-18 in the astrocytes supernatant, while the presence of Mito-TEMPO or z-YVAD-FMK or Z-VAD-FMK or NLRP3 bp abolishes most of the cytokine released in the cell medium (Figures 5A,B). However, the results presented in Figure 5 illustrate that while both Z-YVAD-FMK and z-VAD-FMK were more efficient than NLRP3 bp or Mito-TEMPO in inhibiting IL-18 in all the treatments, the caspase-1 inhibitor did not completely abolish the release of IL-1β induced by ATP or LPS or ethanol, suggesting that this cytokine may be produced by other mechanisms (Hanamsagar et al., 2012). No significant variations in the levels of IL-1β or IL-18 were noted in the TLR4-KO astrocytes treated with ATP, LPS or even ethanol (Figures 5A,B).
