Abstract
It has been demonstrated that the combination of three botanical factors of (+)-catechin, baicalin and β-caryophyllene, which exhibit anti-inflammatory effects, with comparatively lower concentrations of each factor, demonstrating a potent synergistic-suppressive effect on the growth of mouse macrophage RAW264.7 cells in vitro, and suggesting it may function as a pharmacologic tool for managing inflammatory diseases. The present study was undertaken to determine the suppressive effects of (+)-catechin, baicalin or β-caryophyllene on the production of inflammatory cytokines, including TNF-α, IL-6 and IL-1β, which was enhanced by lipopolysaccharide (LPS) in RAW264.7 cells in vitro. The cells were cultured for 3 days without botanical factors, followed by incubation for 5 h in the presence of either vehicle, (+)-catechin [1 µg/ml (3.45 µM)], baicalin [1 µg/ml (2.24 µM)], or β-caryophyllene [1 µg/ml (5 µM)] with or without LPS (100 ng/ml); this did not have significant effects on the number of RAW264.7 cells. The production of TNF-α, IL-6 and IL-1β was not altered by the addition of (+)-catechin, baicalin, β-caryophyllene, or the three combined factors in RAW264.7 cells without LPS. LPS treatment caused a marked production of TNF-α, IL-6, and IL-1β. This enhancement was suppressed by the addition of (+)-catechin, baicalin or β-caryophyllene. Of note, the production of these cytokines was additively suppressed by the combination of the three factors in macrophages. Thus, the combination of (+)-catechin, baicalin and β-caryophyllene was found to reveal a potent suppressive effect on cytokine production in macrophages in vitro. This composition may be a useful tool as a potent anti-inflammatory agent.