A second enzyme protecting mineralocorticoid receptors from glucocorticoid occupancy.
We have confirmed that A6 cells (derived from kidney of Xenopus laevis), which contain both mineralocorticoid and glucocorticoid receptors, do not normally possess 11 beta-hydroxysteroid dehydroxgenase (11 beta-HSD1 or 11 beta-HSD2) enzymatic activity and so are without apparent "protective" enzymes. A6 cells do not convert the glucocorticoid corticosterone to 11-dehydrocorticosterone but do, however, possess steroid 6 beta-hydroxylase that transforms corticosterone to 6 beta-hydroxycorticosterone. This hydroxylase is cytochrome P-450 3A (CYP3A). We have now determined the effects of 3 alpha,5 beta-tetrahydroprogesterone and chenodeoxycholic acid (both inhibitors of 11 beta-HSD1) and 11-dehydrocorticosterone and 11 beta-hydroxy-3 alpha,5 beta-tetrahydroprogesterone (inhibitors of 11 beta-HSD2) and carbenoxalone, which inhibits both 11 beta-HSD1 and 11 beta-HSD2, on the actions and metabolism of corticosterone and active Na+ transport [short-circuit current (Isc)] in A6 cells. All of these 11 beta-HSD inhibitory substances induced a significant increment in corticosterone-induced Isc, which was detectable within 2 h. However, none of these agents caused an increase in Isc when incubated by themselves with A6 cells. In all cases, the additional Isc was inhibited by the mineralocorticoid receptor (MR) antagonist, RU-28318, whereas the original Isc elicited by corticosterone alone was inhibited by the glucocorticoid receptor antagonist, RU-38486. In separate experiments, each agent was shown to significantly inhibit metabolism of corticosterone to 6 beta-hydroxycorticosterone in A6 cells, and a linear relationship existed between 6 beta-hydroxylase inhibition and the MR-mediated increase in Isc in the one inhibitor tested. Troleandomycin, a selective inhibitor of CYP3A, inhibited 6 beta-hydroxylase and also significantly enhanced corticosterone-induced Isc at 2 h. These experiments indicate that the enhanced MR-mediated Isc in A6 cells may be related to inhibition of 6 beta-hydroxylase activity in these cells and that this 6 beta-hydroxylase (CYP3A) may be protecting the expression of corticosterone-induced active Na+ transport in A6 cells by MR-mediated mechanism(s).
Morris
D J
DJ
Department of Pathology, Miriam Hospital, Lifespan, Rhode Island, USA.
Latif
S A
SA
Rokaw
M D
MD
Watlington
C O
CO
Johnson
J P
JP
D042842:11-beta-Hydroxysteroid Dehydrogenases; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D002460:Cell Line / Q000187:drug effects / Q000502:physiology; D003345:Corticosterone / Q000494:pharmacology; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism / Q000502:physiology*; D004553:Electric Conductivity; D005938:Glucocorticoids / Q000378:metabolism*; D006913:Hydroxysteroid Dehydrogenases / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D018942:Macrolides / Q000494:pharmacology; D000451:Mineralocorticoid Receptor Antagonists; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors / Q000502:physiology*; D011965:Receptors, Glucocorticoid / Q000037:antagonists & inhibitors; D018161:Receptors, Mineralocorticoid / Q000378:metabolism*; D013148:Spironolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D013250:Steroid Hydroxylases / Q000378:metabolism; D014982:Xenopus laevis
D065607:Cytochrome P-450 Enzyme Inhibitors; D005938:Glucocorticoids; D007527:Isoenzymes; D018942:Macrolides; D000451:Mineralocorticoid Receptor Antagonists; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; D013148:Spironolactone; C041006:RU 28318; D003577:Cytochrome P-450 Enzyme System; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; C022657:steroid hormone 6-beta-hydroxylase; D010089:Oxidoreductases, N-Demethylating; D003345:Corticosterone