REGULATION OF PROSTAGLANDIN-H SYNTHASE-2 EXPRESSION IN HUMAN MONOCYTES BY THE MARINE NATURAL-PRODUCTS MANOALIDE AND SCALARADIAL - NOVEL EFFECTS INDEPENDENT OF INHIBITION OF LIPID MEDIATOR PRODUCTION

The marine natural products manoalide and scalaradial are potent anti-inflammatory agents that inactivate the enzyme phospholipase A(2) (PLA(2)) in vitro. To study the mechanism of inhibition of prostaglandin E(2) (PGE(2)) production in human monocytes by manoalide and scalaradial, lipopolysaccharide (LPS)-induced prostaglandin biosynthesis and induction of prostaglandin H synthase (PGHS) were evaluated. LPS (10 ng/mL) and interleukin-1 beta (IL-1 beta, 50-1000 ng/mL) but not tumor necrosis factor alpha (TNF alpha, 300 ng/mL) induced the expression of the PGHS-2 isoform as determined by immunoblot analysis with a specific polyclonal antibody for PGHS-2. Manoalide and scalaradial (1-10 mu M) inhibited LPS-induced endogeneous PGE(2) production, reduced the LPS-induced PGHS activity, and reduced the expression of PGHS-2. Indomethacin [a PGHS inhibitor (0.01 to 0.1 mu M)], zileuton [a 5-lipoxygenase inhibitor (3-10 mu M)], and WEB-2806 [a platelet-activating factor (PAF) antagonist (30 mu M)] did not affect the LPS-induced expression of PGHS-2 in human monocytes. These results suggest that modulation of lipid mediator production by manoalide or scalaradial may not be involved in the observed effects on the expression of PGHS-2 Manoalide and scalaradial also inhibited the release of IL-1 beta and TNF alpha from LPS-stimulated monocytes. Expression of PGHS-2 induced by either LPS or IL-1 beta was blocked by the IL-1 receptor antagonist (IL-1ra, 2 mu g/mL) but not by rolipram, a phosphodiesterase IV inhibitor that inhibits TNF alpha but not IL-1 beta release. Similar to LPS, IL-1 beta-induced PGHS-2 expression was apparently not regulated by lipid mediators such as prostaglandins, leukotrienes or PAF as determined with specific inhibitors and antagonists. Scalaradial and to some extent manoalide were capable of blocking the IL-1 beta-induced expression of PGHS-2. These results indicate that IL-1 beta is the predominant cytokine responsible for the induction of PGHS-2 in me human monocyte. Furthermore, marine natural products such as scalaradial have novel effects on the IL-1 beta-mediated induction of PGHS-2 in human monocytes, which appears to be independent of effects on lipid mediator production.