Effects of HSM on ATP-mediated caspase-1 gene expression and activation in human macrophages.
Cells were pretreated with HSM extracts (1 or 2%) for 20 h, followed by treatment with LPS (0.5 μg/ml) for 3 h and subsequently ATP (5 mM) for 1 h. (a) The mRNA expression levels of caspase-1 were determined by RT-PCR analysis. (b) Caspase-1 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) The secretion of caspase-1 subunit p20 into the supernatants of THP-1 macrophages was assessed by ELISA. (d) Cell lysates and culture supernatants were Western-blotted to detect pro-caspase-1 p45 and caspase-1 subunit p20. Data are presented as means ± SE of three experiments preformed in duplicate. #P < 0.01 versus untreated cells. *P < 0.01 versus HSM-untreated control (ethanol) cells. †P < 0.05 versus HSM (1%) treated cells.