Effects of HSM on inflammasome components and caspase-4 activation in human macrophages.
Cells were pretreated with various concentrations (1 or 2%) of HSM extract for 20 h, followed by treatment with LPS (0.5 μg/ml) for 3 h and ATP (5 mM) for 1 h. (a) The mRNA expression levels of ASC, NLRP1, NLRP3 and caspase-4 were determined by RT-PCR, using β-actin as the internal control. (b) ASC, NLRP1, NLRP3 and caspase-4 mRNAs were quantified using real-time PCR. β-actin gene expression was used for normalization. The results are expressed as fold changes, considering one as the value of untreated cells. (c) Cell lysates were analyzed by Western blot analysis using specific anti-ASC, anti-NLRP1 and anti-NLRP3 antibodies. (d) Cell lysates were analyzed for protein levels of caspase-4 by Western blot analysis. β-actin was used as an internal control. Data are presented as means ± SE of three experiments preformed in duplicate. #P < 0.01 versus untreated cells. versus HSM-untreated control (ethanol) cells. *P < 0.01 versus HSM-untreated control (ethanol) cells.