Stimulation of arachidonic acid metabolism in primary cultures of osteoblast-like cells by hormones and drugs
The effects of parathyroid hormone (PTH), dihydroxycholecalciferol (1,25-(OH)2 D3), 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 (/sup 14/C)-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 (/sup 3/H) standard prostanoids. Osteoblasts showed a basal release of the prostanoids 6-keto-PGF1 alpha, TXB2, PGF2 alpha, PGE2, PGD2 and PGB2, 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)2D3 was found to slightly inhibit the prostanoid release. These results indicate: (1) primary cultures of osteoblasts synthesize several prostaglandins, thromboxane B2 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)2D3.
- Research Organization:
- Univ. of Leiden, The Netherlands
- OSTI ID:
- 5655793
- Journal Information:
- Prostaglandins; (United States), Vol. 28:6
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ARACHIDONIC ACID
METABOLISM
PROSTAGLANDINS
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CELL CULTURES
CHICKENS
CHOLECALCIFEROL
EMBRYOS
LIQUID COLUMN CHROMATOGRAPHY
METABOLITES
PARATHORMONE
RESPONSE MODIFYING FACTORS
THROMBIN
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
BIRDS
CARBOXYLIC ACIDS
CHROMATOGRAPHY
COAGULANTS
DRUGS
ENZYMES
FOWL
HEMATOLOGIC AGENTS
HEMOSTATICS
HORMONES
HYDROLASES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDE HORMONES
PEPTIDE HYDROLASES
SEPARATION PROCESSES
SERINE PROTEINASES
SYNTHESIS
VERTEBRATES
VITAMIN D
VITAMINS
550201* - Biochemistry- Tracer Techniques