Study of the Modulation of Cytokine Release by Natural Compounds with Pharmacological Properties Using Cell-Based Systems

Background: The screening of the pharmacological properties of natural compounds (i.e., anti-inflammatory effects) may take advantage of some specific cell-based systems. Parthenolide (PTN) and Copaifera langsdorfii (Copaiba) are natural compounds used to prevent and treat headache and migraine and in inflammatory diseases involving respiratory airways, genital-urinary apparatus and skin, respectively, but their effects at the cellular level are poorly understood. Methods: Mouse BV-2 microglia and human THP-1 monocyte cell lines were used. The nuclear translocation of nuclear factor (NF)-kB was evaluated by Western blotting analysis. The secretion of inflammatory cytokines (interleukin (IL)-1\u3b2, IL-6, tumor necrosis factor-\u3b1 (TNF\u3b1)) was evaluated by immunometric assays (ELISA). Results: Treatment of BV-2 cells with 1 \u3bcM PTN and of THP-1 cells with 10 \u3bcM Copaiba oleoresin (OR), containing diterpene acids, diterpenes and sesquiterpenes, strongly reduced the NF-kB translocation to the cell nucleus induced by 1 \u3bcg/mL lipopolysaccaride (LPS). In BV-2 cells, PTN reduced IL-6 secretion in a dosedependent manner (-29% at 200 nM, P < 0.001; -45% at 1 \u3bcM, P < 0.001; -98% at 5 \u3bcM, P < 0.001; ANOVA). Moreover, at 5 \u3bcm (highest concentration tested) PTN also reduced TNF-\u3b1 secretion (-54%, P < 0.001). Preincubation of LPS-stimulated THP-1 monocytes with OR (dose-range: 0.1-10 mM), reduced the release of all tested cytokines (IL-1\u3b2, IL-6, TNF-\u3b1). Conclusions: The results obtained provide strong evidence that both cell-based models are useful to validate the anti-inflammatory properties of PTN and OR at the cellular level and suggest that they are related to inhibition of cytokine secretion and NF-\u3baB nuclear translocation