Effects of HSM on IL-1β gene expression and secretion in human macrophages.
Cells were pre-treated with either 1 or 2% of HSM extract for 20 h, followed by treatment with LPS (0.5 μg/ml) for 3 h and with ATP (5 mM) for 1 h. (a) The mRNA expression levels of IL-1β were determined by RT-PCR analysis. (b) IL-1β 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 amount of IL-1β in cell culture supernatants was detected by ELISA. (d) The presence of IL-1β in cell lysates and cell culture supernatants were analyzed by Western blot analysis. Data are presented as means ± SE of three experiments performed in duplicate. #P < 0.01 versus untreated cells. *P < 0.01 versus HSM-untreated (ethanol-treated) control cells. †P < 0.05 versus HSM (1%) treated cells.