Suppression of cultured bovine adrenocortical zona glomerulosa cell aldosterone synthesis by steroids and its prevention by antioxidants

J. F. Crivello, Peter J Hornsby, G. N. Gill

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Previous work suggested that autoregulation of adrenocortical function by steroids may be important in the development of different functional zones of the adrenal cortex, possibly through the generation of oxygen-derived free radicals after an interaction with cytochrome P-450s. The present experiments investigate the effects of pretreatment with various steroids on subsequent conversion of deoxycorticosterone (DOC) to aldosterone by primary cultures of bovine adrenocortical zona glomerulosa cells. The products of DOC metabolism, quantitated by high performance liquid chromatography, were aldosterone, 18-hydroxycorticosterone (18-OH B), 18-hydroxydeoxycorticosterone (18-OH DOC), and corticosterone. Corticosterone was the major product formed (approximately 80%), whereas 18-OH B and 18-OH DOC were formed in equal amounts (approximately 10%) and aldosterone was formed in lesser amounts (1%). The maximal rate of production of aldosterone using DOC as precursor was twice the maximal rate using corticosterone as precursor. After plating of glomerulosa cells in culture, conversion of DOC to aldosterone and 18-OH corticosterone decreased with apparent first order kinetics (t( 1/2 ) = 10 and 17 h, respectively) over a time period in which DOC conversion to corticosterone declined only slightly (t( 1/2 ) = 50 h). Addition of 100 μM butylated hydroxyanisole, 50 nM H2SeO3, and 100 mM dimethyl sulfoxide to the culture medium slowed the loss of formation of aldosterone, 18-OH B, and corticosterone but did not halt it (t( 1/2 ) = 23, 26, and 65 h, respectively). In order of potency, pretreatment with androstenedione, cortisol, 18-OH DOC, and 11β-hydroxyandrostenedione accelerated the loss of the subsequent conversion of DOC to aldosterone and 18-OH B. Aldosterone production decreased at a faster rate than production of corticosterone or 18-OH B. Pretreatment with cortisol also depressed the subsequent conversion of corticosterone to aldosterone. Analysis of effective steroid structures suggested interaction with glomerulosa cytochrome P-450(11β) as a requirement for activity. Antioxidants prevented or reduced the effects of steroids on DOC metabolism. The most potent agent was the OH.-scavenger dimethyl sulfoxide, butylated hydroxyanisole was also effective, whereas the natural antioxidant α-tocopherol was not. The suppression of corticosterone conversion to aldosterone by cortisol makes it unlikely that the reduction in glomerulosa cytochrome P-450(11β) activity (i.e. reduction in the production of the precursor corticosterone) is responsible for the reduction in DOC conversion to aldosterone. It is more likely that glomerulosa aldosterone production is reduced by the interaction of corticol with glomerulosa cytochrome P-450(11β) with the concomitant production of oxygen-derived free radicals resulting in the initiation of lipid peroxidation and formation of lipid hydroperoxides. Lipid hydroperoxides then cause loss of the terminal cytochrome P-450 (P-450(CMO), corticosterone methyl oxidase) in the aldosterone biosynthetic pathway at a faster rate than that of glomerulosa cytochrome P-450(11β).

Original languageEnglish (US)
Pages (from-to)235-242
Number of pages8
JournalEndocrinology
Volume113
Issue number1
StatePublished - 1983
Externally publishedYes

Fingerprint

Zona Glomerulosa
Aldosterone
Corticosterone
Desoxycorticosterone
Antioxidants
Steroids
Cytochrome P-450 Enzyme System
Butylated Hydroxyanisole
Hydrocortisone
Lipid Peroxides
Dimethyl Sulfoxide
Free Radicals
18-Hydroxycorticosterone
Oxygen
Tocopherols
Androstenedione
Adrenal Cortex
Biosynthetic Pathways
Cytochromes
Lipid Peroxidation

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Suppression of cultured bovine adrenocortical zona glomerulosa cell aldosterone synthesis by steroids and its prevention by antioxidants. / Crivello, J. F.; Hornsby, Peter J; Gill, G. N.

In: Endocrinology, Vol. 113, No. 1, 1983, p. 235-242.

Research output: Contribution to journalArticle

@article{887dbdb597814bac8e5edf092b3d9c27,
title = "Suppression of cultured bovine adrenocortical zona glomerulosa cell aldosterone synthesis by steroids and its prevention by antioxidants",
abstract = "Previous work suggested that autoregulation of adrenocortical function by steroids may be important in the development of different functional zones of the adrenal cortex, possibly through the generation of oxygen-derived free radicals after an interaction with cytochrome P-450s. The present experiments investigate the effects of pretreatment with various steroids on subsequent conversion of deoxycorticosterone (DOC) to aldosterone by primary cultures of bovine adrenocortical zona glomerulosa cells. The products of DOC metabolism, quantitated by high performance liquid chromatography, were aldosterone, 18-hydroxycorticosterone (18-OH B), 18-hydroxydeoxycorticosterone (18-OH DOC), and corticosterone. Corticosterone was the major product formed (approximately 80{\%}), whereas 18-OH B and 18-OH DOC were formed in equal amounts (approximately 10{\%}) and aldosterone was formed in lesser amounts (1{\%}). The maximal rate of production of aldosterone using DOC as precursor was twice the maximal rate using corticosterone as precursor. After plating of glomerulosa cells in culture, conversion of DOC to aldosterone and 18-OH corticosterone decreased with apparent first order kinetics (t( 1/2 ) = 10 and 17 h, respectively) over a time period in which DOC conversion to corticosterone declined only slightly (t( 1/2 ) = 50 h). Addition of 100 μM butylated hydroxyanisole, 50 nM H2SeO3, and 100 mM dimethyl sulfoxide to the culture medium slowed the loss of formation of aldosterone, 18-OH B, and corticosterone but did not halt it (t( 1/2 ) = 23, 26, and 65 h, respectively). In order of potency, pretreatment with androstenedione, cortisol, 18-OH DOC, and 11β-hydroxyandrostenedione accelerated the loss of the subsequent conversion of DOC to aldosterone and 18-OH B. Aldosterone production decreased at a faster rate than production of corticosterone or 18-OH B. Pretreatment with cortisol also depressed the subsequent conversion of corticosterone to aldosterone. Analysis of effective steroid structures suggested interaction with glomerulosa cytochrome P-450(11β) as a requirement for activity. Antioxidants prevented or reduced the effects of steroids on DOC metabolism. The most potent agent was the OH.-scavenger dimethyl sulfoxide, butylated hydroxyanisole was also effective, whereas the natural antioxidant α-tocopherol was not. The suppression of corticosterone conversion to aldosterone by cortisol makes it unlikely that the reduction in glomerulosa cytochrome P-450(11β) activity (i.e. reduction in the production of the precursor corticosterone) is responsible for the reduction in DOC conversion to aldosterone. It is more likely that glomerulosa aldosterone production is reduced by the interaction of corticol with glomerulosa cytochrome P-450(11β) with the concomitant production of oxygen-derived free radicals resulting in the initiation of lipid peroxidation and formation of lipid hydroperoxides. Lipid hydroperoxides then cause loss of the terminal cytochrome P-450 (P-450(CMO), corticosterone methyl oxidase) in the aldosterone biosynthetic pathway at a faster rate than that of glomerulosa cytochrome P-450(11β).",
author = "Crivello, {J. F.} and Hornsby, {Peter J} and Gill, {G. N.}",
year = "1983",
language = "English (US)",
volume = "113",
pages = "235--242",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Suppression of cultured bovine adrenocortical zona glomerulosa cell aldosterone synthesis by steroids and its prevention by antioxidants

AU - Crivello, J. F.

AU - Hornsby, Peter J

AU - Gill, G. N.

PY - 1983

Y1 - 1983

N2 - Previous work suggested that autoregulation of adrenocortical function by steroids may be important in the development of different functional zones of the adrenal cortex, possibly through the generation of oxygen-derived free radicals after an interaction with cytochrome P-450s. The present experiments investigate the effects of pretreatment with various steroids on subsequent conversion of deoxycorticosterone (DOC) to aldosterone by primary cultures of bovine adrenocortical zona glomerulosa cells. The products of DOC metabolism, quantitated by high performance liquid chromatography, were aldosterone, 18-hydroxycorticosterone (18-OH B), 18-hydroxydeoxycorticosterone (18-OH DOC), and corticosterone. Corticosterone was the major product formed (approximately 80%), whereas 18-OH B and 18-OH DOC were formed in equal amounts (approximately 10%) and aldosterone was formed in lesser amounts (1%). The maximal rate of production of aldosterone using DOC as precursor was twice the maximal rate using corticosterone as precursor. After plating of glomerulosa cells in culture, conversion of DOC to aldosterone and 18-OH corticosterone decreased with apparent first order kinetics (t( 1/2 ) = 10 and 17 h, respectively) over a time period in which DOC conversion to corticosterone declined only slightly (t( 1/2 ) = 50 h). Addition of 100 μM butylated hydroxyanisole, 50 nM H2SeO3, and 100 mM dimethyl sulfoxide to the culture medium slowed the loss of formation of aldosterone, 18-OH B, and corticosterone but did not halt it (t( 1/2 ) = 23, 26, and 65 h, respectively). In order of potency, pretreatment with androstenedione, cortisol, 18-OH DOC, and 11β-hydroxyandrostenedione accelerated the loss of the subsequent conversion of DOC to aldosterone and 18-OH B. Aldosterone production decreased at a faster rate than production of corticosterone or 18-OH B. Pretreatment with cortisol also depressed the subsequent conversion of corticosterone to aldosterone. Analysis of effective steroid structures suggested interaction with glomerulosa cytochrome P-450(11β) as a requirement for activity. Antioxidants prevented or reduced the effects of steroids on DOC metabolism. The most potent agent was the OH.-scavenger dimethyl sulfoxide, butylated hydroxyanisole was also effective, whereas the natural antioxidant α-tocopherol was not. The suppression of corticosterone conversion to aldosterone by cortisol makes it unlikely that the reduction in glomerulosa cytochrome P-450(11β) activity (i.e. reduction in the production of the precursor corticosterone) is responsible for the reduction in DOC conversion to aldosterone. It is more likely that glomerulosa aldosterone production is reduced by the interaction of corticol with glomerulosa cytochrome P-450(11β) with the concomitant production of oxygen-derived free radicals resulting in the initiation of lipid peroxidation and formation of lipid hydroperoxides. Lipid hydroperoxides then cause loss of the terminal cytochrome P-450 (P-450(CMO), corticosterone methyl oxidase) in the aldosterone biosynthetic pathway at a faster rate than that of glomerulosa cytochrome P-450(11β).

AB - Previous work suggested that autoregulation of adrenocortical function by steroids may be important in the development of different functional zones of the adrenal cortex, possibly through the generation of oxygen-derived free radicals after an interaction with cytochrome P-450s. The present experiments investigate the effects of pretreatment with various steroids on subsequent conversion of deoxycorticosterone (DOC) to aldosterone by primary cultures of bovine adrenocortical zona glomerulosa cells. The products of DOC metabolism, quantitated by high performance liquid chromatography, were aldosterone, 18-hydroxycorticosterone (18-OH B), 18-hydroxydeoxycorticosterone (18-OH DOC), and corticosterone. Corticosterone was the major product formed (approximately 80%), whereas 18-OH B and 18-OH DOC were formed in equal amounts (approximately 10%) and aldosterone was formed in lesser amounts (1%). The maximal rate of production of aldosterone using DOC as precursor was twice the maximal rate using corticosterone as precursor. After plating of glomerulosa cells in culture, conversion of DOC to aldosterone and 18-OH corticosterone decreased with apparent first order kinetics (t( 1/2 ) = 10 and 17 h, respectively) over a time period in which DOC conversion to corticosterone declined only slightly (t( 1/2 ) = 50 h). Addition of 100 μM butylated hydroxyanisole, 50 nM H2SeO3, and 100 mM dimethyl sulfoxide to the culture medium slowed the loss of formation of aldosterone, 18-OH B, and corticosterone but did not halt it (t( 1/2 ) = 23, 26, and 65 h, respectively). In order of potency, pretreatment with androstenedione, cortisol, 18-OH DOC, and 11β-hydroxyandrostenedione accelerated the loss of the subsequent conversion of DOC to aldosterone and 18-OH B. Aldosterone production decreased at a faster rate than production of corticosterone or 18-OH B. Pretreatment with cortisol also depressed the subsequent conversion of corticosterone to aldosterone. Analysis of effective steroid structures suggested interaction with glomerulosa cytochrome P-450(11β) as a requirement for activity. Antioxidants prevented or reduced the effects of steroids on DOC metabolism. The most potent agent was the OH.-scavenger dimethyl sulfoxide, butylated hydroxyanisole was also effective, whereas the natural antioxidant α-tocopherol was not. The suppression of corticosterone conversion to aldosterone by cortisol makes it unlikely that the reduction in glomerulosa cytochrome P-450(11β) activity (i.e. reduction in the production of the precursor corticosterone) is responsible for the reduction in DOC conversion to aldosterone. It is more likely that glomerulosa aldosterone production is reduced by the interaction of corticol with glomerulosa cytochrome P-450(11β) with the concomitant production of oxygen-derived free radicals resulting in the initiation of lipid peroxidation and formation of lipid hydroperoxides. Lipid hydroperoxides then cause loss of the terminal cytochrome P-450 (P-450(CMO), corticosterone methyl oxidase) in the aldosterone biosynthetic pathway at a faster rate than that of glomerulosa cytochrome P-450(11β).

UR - http://www.scopus.com/inward/record.url?scp=0020559773&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020559773&partnerID=8YFLogxK

M3 - Article

C2 - 6861699

AN - SCOPUS:0020559773

VL - 113

SP - 235

EP - 242

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 1

ER -