Modulation of interleukin 1β production in macrophages stimulated with lipopolysaccharide by the protein kinase inhibitor staurosporine

Abstract

Inhibitors of key components of intracellular signaling pathways were used to detect differences in the regulation of the production of interleukin 1β (IL-1β) and tumor necrosis factor α (TNFα) in macrophages activated with lipopolysaccharide (LPS). The protein kinase inhibitor staurosporine caused an increase in the level of IL-1β in supernatants of macrophages stimulated with LPS. Calphostin C and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H7) dihydrochloride, specific inhibitors of protein kinase C (PKC), also promoted enhancement of IL-1β secretion, but the effect was not as pronounced as that of staurosporine. In contrast, all three substances inhibited TNFα production. Measurement of IL-1β in lysates and supernatants of macrophage cultures indicated that staurosporine effected enhancement of the production of the cytokine in the cellular fraction, the greater portion of which was not secreted. Kinetics of accumulation of IL-1β mRNA and production of the cytokine during a 24 h period showed that enhanced production of IL-1β obtained 24 h after LPS stimulation of macrophages in the presence of staurosporine paralleled the increased levels in mRNA specific for the cytokine.