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-kappa B (p65), which is a pivotal regulator of the cellular immune response. Since NF-kappa B is involved in the transcriptional control of several pro-inflammatory and regulatory genes, we investigated the effects of one bifunctional NF-kappa B (p65) inhibiting and two monofunctional NF-kappa B (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 11 alpha,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-kappa B alpha, bcl-2, and cyclin D1. The bifunctional NF-kappa B 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-kappa B (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-kappa B inhibition, but must be coupled to other mechanisms.
Gertsch
Jürg
J
Institute for Pharmaceutical Sciences, Swiss Federal Institute of Technology Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Sticher
Otto
O
Schmidt
Thomas
T
Heilmann
Jörg
J
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D005912:Gliotoxin / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D012717:Sesquiterpenes / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D045789:Sesquiterpenes, Guaiane; D014158:Transcription, Genetic / Q000187:drug effects* / Q000502:physiology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016328:NF-kappa B; D012333:RNA, Messenger; D012717:Sesquiterpenes; D045789:Sesquiterpenes, Guaiane; C001329:helenalin; D005912:Gliotoxin