Suppression of the NF-kappa B signalling pathway by ergolide, sesquiterpene lactone, in HeLa cells

Abstract

We have previously reported that ergolide, a sesquiterpene lactone isolated from Inula britannica, suppresses inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by inhibiting nuclear factor-kappa B (NF-kappa B) in RAW 264.7 macrophages. In this study, we show that ergolide suppresses the DNA binding activity of NF-kappa B and nuclear translocation of NF-kappa B p65 subunit, leading to the inhibition of NF-kappa B-dependent gene transcription in 12-O-tetradecanoylphorbol 13-acetate (TPA)-stimulated HeLa cells. We also show that ergolide decreases the degradation and phosphorylation of I kappa B, an inhibitory protein of NF-kappa B, and this effect is accompanied by a simultaneous reduction of I kappa B kinase (IKK) activity. However, ergolide does not inhibit in-vitro IKK activity directly, suggesting the possible involvement of upstream IKK kinases in the regulation of NF-kappa B activation. Furthermore, ergolide-mediated protein kinase C alpha (PKC alpha) inhibition is involved in reduction of NF-kappa B inhibition, as demonstrated by the observation that dominant negative PKC alpha, but not p44/42 MAPK and p38 MAPK, inhibits TPA-stimulated reporter gene expression. Taken together, our results suggest that ergolide suppresses NF-kappa B activation through the inhibition of PKC alpha-IKK activity, providing insight for PKC alpha as a molecular target for anti-inflammatory drugs.