The triterpenoid quinonemethide pristimerin inhibits induction of inducible nitric oxide synthase in murine macrophages.
Inducible nitric oxide synthase dependent production of nitric oxide (NO) plays an important role in inflammation. We investigated whether pristimerin ((20alpha)-3-hydroxy-2-oxo-24-nor-friedela-1(10),3,5,7-te traen-carboxylic acid-(29)-methylester), an antitumoral, antimicrobial as well as anti-inflammatory plant compound, has an effect on the inducible NO synthase system in lipopolysaccharide-activated RAW 264.7 macrophages. Pristimerin dose dependently (IC50: 0.2-0.3 microM) reduces nitrite accumulation, a parameter for NO synthesis, in supernatants of lipopolysaccharide-stimulated (1 microg/ml, 20 h) macrophages. This effect correlates with a reduced inducible NO synthase enzyme activity measured by conversion of [3H]L-arginine to [3H]L-citrulline and significantly lower levels of enzyme protein (Western blotting) in homogenates of cells cotreated with lipopolysaccharide and pristimerin (12 h). Northern blot analysis and polymerase chain reaction (PCR) showed decreased inducible NO synthase mRNA levels in activated macrophages exposed to pristimerin (4 h). Electrophoretic mobility shift assay (EMSA) demonstrated a markedly reduced binding activity of nuclear factor-kappa B (NFkappaB) in nuclear extracts of pristimerin-treated cells. These results suggest that pristimerin inhibits the induction of inducible NO synthase by a mechanism which involves inhibition of NFkappaB activation. This feature of pristimerin is likely to contribute to its anti-inflammatory activity.
Dirsch
V M
VM
Institute of Pharmacology, Toxicology and Pharmacy, Ludwig-Maximilians University, Munich, Germany. dirsch@pharmtox.vetmed.uni-muenchen.de
Kiemer
A K
AK
Wagner
H
H
Vollmar
A M
AM
D002460:Cell Line; D051546:Cyclooxygenase 2; D004790:Enzyme Induction / Q000187:drug effects; D007527:Isoenzymes / Q000096:biosynthesis; D008070:Lipopolysaccharides; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D053978:Pentacyclic Triterpenes; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014315:Triterpenes / Q000494:pharmacology*; D019323:omega-N-Methylarginine / Q000494:pharmacology
D007527:Isoenzymes; D008070:Lipopolysaccharides; D016328:NF-kappa B; D053978:Pentacyclic Triterpenes; D011759:Pyrrolidines; D013859:Thiocarbamates; D014315:Triterpenes; C020972:pyrrolidine dithiocarbamic acid; D019323:omega-N-Methylarginine; D019001:Nitric Oxide Synthase; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C050414:celastrol