To gain further insight into how ethanol affects inflammasome activation and which type of inflammasome was activated in astroglial cells, we used highly enriched astrocytes in primary culture, and the gene expression of different inflammasomes were assessed. An RT-PCR analysis revealed (Figure 2A) that although inflammasomes NLRP3, AIM2, NLRP1, and IPAF are expressed in astrocytes; the NLRP3 mRNA level was higher than the other inflammasomes, and the ethanol treatment (10 and 50 mM) was capable of up-regulating the NLRP3 mRNA levels (Figure 2B). The immunofluorescence studies also revealed that the NLRP3 protein expression is expressed in the cytosol of GFAP+ cells (Figure 2C), although higher NLRP3 expression are noted in WT astrocytes treated with LPS, ATP or ethanol (10, 50 mM).
