9-(tetrahydro-2-furyl)-adenine and chelerythrine

Inhibitors of intracellular signalling events, including enzyme inhibitors, are often used to investigate signal transduction pathways. We examined whether some inhibitors that act on the ATP site of enzymes are also potent adenosine receptor antagonists. Competitive radioligand binding assays in membranes or brain sections show that genistein, chelerythrine, and SQ22536 [9-(tetrahydro-2'-furyl) adenine] block A(1), A(2A), and A(3) adenosine receptors in concentrations of these drugs commonly used to examine cellular signalling (K(i) of [(3)H]-DPCPX (1,3-dipropyl-8-cyclopentylxanthine) competition mean (95% confidence interval): 2.6 (1.5-4.8) microM, 5.7 (2.1-15.8) microM, 59.4 (17.3-203.8) microM; K(i) of [(3)H]-SCH58261 [5-amino-7-(2-phenylethyl)-2-(2-furyl)-pyrazolo[4,3-e]-1,2,4-triazolo[1,5-c]pyrimidine] competition: 15.3 (8.1-28.8) microM, 37.6 (10.3-137.4) microM, 16.7 (11.5-24.3) microM for genistein, chelerythrine, and SQ22536, respectively). Given that adenosine receptors are present on most cells, that adenosine is often present, and that adenosine receptors interact functionally with several signalling pathways, these results may be of significance also when studying signalling via other receptors.

Topics: Adenine; Adenylyl Cyclase Inhibitors; Alkaloids; Animals; Benzophenanthridines; CHO Cells; Cricetinae; Cyclic AMP Response Element-Binding Protein; Cytoplasm; Dose-Response Relationship, Drug; Enzyme Inhibitors; Genistein; Male; Mice; Mitogen-Activated Protein Kinases; Phenanthridines; Phosphorylation; Protein Kinase C; Purinergic P1 Receptor Antagonists; Pyrimidines; Rats; Rats, Sprague-Dawley; Signal Transduction; Triazoles; Xanthines

To investigate the function and mechanism of fasting on the production of corticosterone in vitro by zona fasciculata-reticularis (ZFR) cells from ovariectomized (OVX) rats.. Female rats were OVX for 4 days before decapitation. Rats were fed or fasted for 1 day before experiment. ZFR cells from fed and fasted rats were incubated with adrenocorticotropic hormone (ACTH), forskolin, 8-bromo-3',5'-cyclic adenosine monophosphate, SQ22536, nifedipine, chelerythrine chloride, trilostane or steroidogenic precursors at 37 degrees C for either 60 or 30 minutes. Corticosterone, pregnenolone concentrations in spent media, and the intracellular adenosine 3',5'-cyclic monophosphate (cAMP) concentration were determined by radioimmunoassay. The effects of fasting in response to ACTH on the protein expressions of steroidogenic acute regulatory protein (StAR) or cytochrome P450 side-chain cleavage enzyme (P450scc) in ZFR cells were determined by Western blot analysis.. The concentration of plasma corticosterone in fasted rats was significantly higher than that in fed rats (P<0.01). One-day fasting significantly increased the responsiveness of ZFR cells to ACTH, forskolin, and precursor-stimulated corticosterone productions and to forskolin-stimulated cAMP accumulation. The corticosterone production was reduced in fasted group when adenylyl cyclase was inhibited by SQ22536. The fasting-enhanced level of corticosterone production in ZFR cells was decreased by the administration of nifedipine but not altered by that of chelerythrine chloride. Fasting significantly increased trilostane-stimulated production of pregnenolone in ZFR cells. The activities of enzymes which converting cholesterol, pregnenolone, progesterone, and deoxycorticosterone to corticosterone and the expressions of StAR in ZFR cells were greater in fasted rats than in fed rats.. This study demonstrated that fasting increased the release of corticosterone and the accumulation of cAMP by rat ZFR cells. The action mediated through enhancing the responsiveness to ACTH stimulation, cAMP cascades and the activity of L-type calcium channels. The activities of steroidogenic enzymes including P450scc, 3beta-hydroxysteroid dehydrogenase, 21-hydroxylase, and 11beta-hydroxylase were all enhanced by the fasting treatment.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Adenine; Adrenocorticotropic Hormone; Alkaloids; Animals; Benzophenanthridines; Cells, Cultured; Cholesterol Side-Chain Cleavage Enzyme; Colforsin; Corticosterone; Culture Media, Conditioned; Dihydrotestosterone; Female; Food Deprivation; Nifedipine; Ovariectomy; Phenanthridines; Phosphoproteins; Pregnenolone; Rats; Rats, Sprague-Dawley; Zona Fasciculata; Zona Reticularis