ABSTRACT
OBJECTIVE AND DESIGN: We have investigated the mechanisms of action of aethiopinone, an anti-inflammatory compound from Salvia aethiopis L. roots. MATERIAL AND SUBJECTS: Human neutrophils from healthy volunteers and murine peritoneal macrophages. Swiss mice were randomly divided into groups of six animals. TREATMENT: Test compounds were applied topically in the mouse ear oedema test. In the air pouch, mice received aethiopinone (0.001-0.5 pmol/pouch or 12.5-50 mg/kg p.o.). METHODS: LTB4 production was assayed in human neutrophils and COX-2 and iNOS activities in murine macrophages. Air pouches were induced subcutaneously in mice and injected with zymosan on the day six. Mouse ear oedema was induced by arachidonic acid. Dunnett's t-test was employed for statistical analysis. RESULTS: We have observed potent inhibitory effects on human neutrophil LTB4 production without effects on COX or NOS activities. Aethiopinone is an in vitro inhibitor of 5-LO from human neutrophils (IC50 = 0.11 microM). In addition, aethiopinone reduced leukocyte accumulation and showed in vivo inhibitory activity on this enzyme. CONCLUSIONS: Our results indicate that inhibition of 5-LO could participate in the anti-inflammatory properties of this natural product.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Lipoxygenase Inhibitors/pharmacology , Naphthoquinones/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Arachidonic Acid , Cyclooxygenase 1 , Cyclooxygenase 2 , Dinoprostone/metabolism , Ear , Edema/chemically induced , Edema/drug therapy , Edema/metabolism , Humans , Inflammation/metabolism , Isoenzymes/metabolism , Leukotriene B4/biosynthesis , Macrophages, Peritoneal/enzymology , Membrane Proteins , Mice , Naphthoquinones/therapeutic use , Neutrophils/enzymology , Neutrophils/metabolism , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type II , Phospholipases A/antagonists & inhibitors , Prostaglandin-Endoperoxide Synthases/metabolismABSTRACT
The inhibitory effect of two neo-clerodane diterpenoids, E-isolinaridial (EI) and its methylketone derivative (EIM), isolated from Linaria saxatilis var. glutinosa, on PLA2 and other enzyme activities involved in the inflammatory process was studied. Both compounds inhibited human synovial sPLA2 in a concentration-dependent manner with IC50 values of 0.20 and 0.49 microM, respectively, similar to scalaradial. Besides, these compounds decreased the cell-free 5-lipoxygenase activity and A23187-induced neutrophil LTB4 biosynthesis. Another function of human neutrophils, such as receptor-mediated degranulation, was also significantly reduced. In contrast, none of the compounds affected superoxide generation in leukocytes, or cyclooxygenase-1, cyclooxygenase-2 and inducible nitric oxide synthase activities in cell-free assays.
Subject(s)
Diterpenes/pharmacology , Enzyme Inhibitors/pharmacology , Lipoxygenase Inhibitors , Phospholipases A/antagonists & inhibitors , Humans , Leukocyte Elastase/metabolism , Leukotriene B4/biosynthesis , Neutrophils/metabolism , Phospholipases A2ABSTRACT
A new inhibitor of human secretory phospholipase A2 (PLA2), cacospongionolide E (4a), has been isolated from the Tyrrhenian sponge Fasciospongia cavernosa. The structure was proposed on the basis of spectroscopic data and by chemical transformations. The absolute configuration of cacospongionolides 2a-4a was established using the modified Mosher's method. Cacospongionolide E was the most potent inhibitor toward human synovial PLA2, showing higher potency than the reference compound manoalide and exerting no signs of toxicity on human neutrophils. It showed high activity in the Artemia salina bioassay and moderate toxicity in the fish (Gambusia affinis) lethality assay.
