Stimulation of arachidonic acid metabolism in primary cultures of osteoblast-like cells by hormones and drugs

Abstract

The effects of parathyroid hormone (PTH), dihydroxycholecalciferol (1,25-(OH) 2 D 3), thrombin, epidermal growth factor (EGF) and 12-o-tetradecanoylphorbol-13-acetate (PMA) on the biosynthesis and release of arachidonic acid metabolites were studied in primary cultures of osteoblast-like cells isolated from 18-day-old chick embryo calvaria. Cells were labelled with ( 14C)-arachidonic acid for 30 h. The radioactive eicosanoids were extracted from the cell culture media after a further 30 h stimulation period and analysed on a PRP-1 column by HPLC. The radioactive products were characterized by co-elution of ( 3H) standard prostanoids. Osteoblasts showed a basal release of the prostanoids 6-keto-PGF 1α, TXB 2, PGF 2α, PGE 2, PGD 2 and PGB 2, the latter being the most abundant one. Indomethacin (10 −5 M) effectively inhibited the basal release, but not that of an as yet unidentified compound. The release of prostanoids was stimulated by PTH (2 U/ml), thrombin (0.4 NIH/ml), EGF (50 ng/ml) and PMA (25 ng/ml), the latter being by far the most potent one. 1,25-(OH) 2D 3 was found to slightly inhibit the prostanoid release. These results indicate: (1) primary cultures of osteoblasts synthesize several prostaglandins, thromboxane B 2 and one unidentified product. (2) the action on bone of PTH and the various drugs tested may be, at least partly, mediated by an increased prostaglandin production by osteoblasts. Clearly this does not apply to 1,25-(OH) 2D 3.