Influence of helenanolide-type sesquiterpene lactones on gene transcription profiles in Jurkat T cells and human peripheral blood cells: anti-inflammatory and cytotoxic effects

Sesquiterpene lactones (SLs) are known to have potent anti-inflammatory and cytotoxic properties. So far, the anti-inflammatory effects have mainly been attributed to their inhibition of DNA-binding of the transcription factor NF-kappaB (p65), which is a pivotal regulator of the cellular immune response. Since NF-kappaB is involved in the transcriptional control of several pro-inflammatory and regulatory genes, we investigated the effects of one bifunctional NF-kappaB (p65) inhibiting and two monofunctional NF-kappaB (p65) inactive helenanolide-type SLs on PMA and LPS-induced mRNA expression in CD4(+) Jurkat T and human peripheral blood mononuclear cells (PBMCs) with reverse transcription real-time PCR (RT-rt-PCR). The monofunctional SLs 11alpha,13-dihydrohelenalin acetate (DHAc) and chamissonolide significantly reduced mitogen-induced cytokine and iNOS mRNA levels in PBMCs and Jurkat T cells at low micromolar concentrations. DHAc also showed significant effects on the gene expression of the house-keeping genes GAP-DH and beta-actin, as well as on NF-ATc, p65, I-kappaBalpha, bcl-2, and cyclin D1. The bifunctional NF-kappaB inhibitor helenalin not only effectively inhibited pro-inflammatory gene expression, but also strongly down-regulated all investigated mRNA levels in a time-dependent manner. Flow cytometry and caspase-8 and -3 assays revealed that helenalin strongly and DHAc moderately induced apoptosis in Jurkat T cells, whereas chamissonolide caused cytoprotective effects. In PBMCs, DHAc and chamissonolide did not inhibit NF-kappaB (p65) DNA-binding at concentrations effective on the transcriptome. Thus, it can be concluded that the biological effects of SLs are not only due to NF-kappaB inhibition, but must be coupled to other mechanisms. (C) 2003 Elsevier Inc. All rights reserved.