Ultralow concentrations of ibuprofen activate cell prostaglandin synthesis

Abstract

The interest in the prostaglandin (PG) synthesis by animal cells today grows steadily because of the difficulties in obtaining them by any other way. Murine peritoneal macrophages can under certain conditions synthesize large amounts of PGs. The effect of well-known nonsteroidal anti-inflammatory drug ibuprofen on PG synthesis by the cells using a high-performance liquid chromatography (HPLC) method with fluorescence detection of 4-bromomethyl-7-methoxy-coumarin (BrMMC) derivatives was studied. In our case, the main metabolites were PGE2 and PGF2a. The PG synthesis activation effect was shown by ibuprofen concentrations in the 10(-10)-10(-14) M range with the maximum effect of 10(-12)M. In this case, the ibuprofen effect was comparable in value with the effect of well-known cell PG synthesis activator--calcium ionophore A23187. Although the exact mechanism of such an effect is not clear at the moment, at low concentration, ibuprofen itself is able to activate PG synthesis in murine peritoneal macrophages.

Prostaglandin E2biphasic control of lymphocyte proliferation: inhibition by picomolar concentrations

“…Splenocytes in 6‐well plates were cultured in complete RPMI 1640 (4×10 6 /ml) and at the chosen time cell supernatants were collected and gently centrifuged. Prostanoids synthesized by the cells were determined as derivatives of 4‐bromomethyl‐7‐methoxycoumarin (BrMMC, Sigma) by HPLC with fluorescent detection as described previously [13]. Briefly, cell supernatants were acidified with 1 M HCl to pH 3.0 prior to extraction with ethyl acetate.…”

Kinetics of prostanoid synthesis by macrophages is regulated by arachidonic acid sources

“…Prostanoids synthesized by cells or by microsomes were determined as derivatives of BrMMC by HPLC with fluorescent detection as described previously [27]. Adherent macrophages in 96‐well plates were cultured in fresh DMEM supplemented with 0.5% BSA for 1 h, then A23187 or AA was added.…”