scholarly journals Chenodeoxycholic Acid and Deoxycholic Acid Inhibit 11β-Hydroxysteroid Dehydrogenase Type 2 and Cause Cortisol-induced Transcriptional Activation of the Mineralocorticoid Receptor

2002 ◽  
Vol 277 (29) ◽  
pp. 26286-26292 ◽  
Author(s):  
Anita T. Stauffer ◽  
Mascha K. Rochat ◽  
Bernhard Dick ◽  
Felix J. Frey ◽  
Alex Odermatt
Keyword(s):  
Chenodeoxycholic Acid ◽  
Transcriptional Activation ◽  
Mineralocorticoid Receptor ◽  
Deoxycholic Acid ◽  
Hydroxysteroid Dehydrogenase

Quantitative determination and tissue distribution of human 11β-hydroxysteroid dehydrogenase, hexose-6-phosphate dehydrogenase, glucose-6-phosphate transporter, glucocorticoid receptor and mineralocorticoid receptor mRNAs

2010 ◽  
Vol 2 (1) ◽  
Author(s):  
Shuji Ohno ◽  
Masanori Ohta ◽  
Shizuo Nakajin
Keyword(s):  
Small Intestine ◽  
Quantitative Determination ◽  
Mineralocorticoid Receptor ◽  
Hydroxysteroid Dehydrogenase ◽  
Phosphate Transporter ◽  
Physiological Effects ◽  
Wide Range

Abstract: Glucocorticoid (GC) concentrations in peripheral tissues are precisely regulated by 11β-hydroxysteroid dehydrogenase (HSD) isozymes. When considering the physiological effects of GC in various tissues, quantitative determination of these isozymes and other components involved in corticosteroid signaling is important and informative. We thus performed comprehensive determination of the expression of these mRNAs in a wide range of human tissues.: An absolute comparison of mRNA expression of human 11β-HSD isozymes, hexose-6-phosphate dehydrogenase (H6PDH), glucose-6-phosphate transporter (G6PT), glucocorticoid receptors (GRs), and mineralocorticoid receptor (MR) was performed by real-time RT-PCR.: Human 11β-HSD type 1 mRNA was strongly expressed in the liver and placenta at comparatively high levels. H6PDH was expressed at low copy number, and comparatively high expression was observed in the kidney, testis, and ovary. G6PT expression was ubiquitous, but marked expression was observed in the liver, kidney, small intestine, and colon. GRα was also ubiquitously expressed at relatively high levels, which were approximately 10-fold higher than those of MR, whereas GRβ levels were below the detection limit in all tissues. 11β-HSD type 2 was predominantly expressed in the kidney, small intestine and colon; however, copy numbers of these transcripts showed a nearly identical pattern to type 1. MR was observed in various tissues examined, but was not fully correlated to the distribution of 11β-HSD type 2.: The present quantitative results were partially consistent with previous studies. This quantification method can thus provide valuable information for understanding the physiological effects and physiological roles of glucocorticoid in humans.

Oxidation of primary bile acids by a 7α-hydroxysteroid dehydrogenase elaborating Clostridium bifermentans soil isolate

1987 ◽  
Vol 33 (8) ◽  
pp. 663-669 ◽  
Author(s):  
J. Derek Sutherland ◽  
C. Noel Williams ◽  
Donna M. Hutchison ◽  
Lillian V. Holdeman
Keyword(s):  
Bile Acids ◽  
Chenodeoxycholic Acid ◽  
Deoxycholic Acid ◽  
Specific Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Ph Optimum ◽  
Reactive Blue 2 ◽  
Cell Extracts ◽  
Initial Drop ◽  
Clostridium Bifermentans

A gram-positive, rod-shaped anaerobe (strain F-6) was isolated from soil. This organism was identified by cellular morphology as well as fermentative and biochemical data as Clostridium bifermentans. Strain F-6 formed 7-ketolithocholic acid from chenodeoxycholic acid and 7-ketodeoxycholic acid from cholic acid in whole cell cultures, but did not transform deoxycholic acid, ursodeoxycholic acid, or ursocholic acid. This reaction is reversible. The structures of 7-ketolithocholic acid and 7-ketodeoxycholic acid were verified by mass spectroscopy and by thin-layer chromatography using Komarowsky's spray reagent. When incubated with the strain F-6 glycine and taurine conjugates of the primary bile acids were partially hydrolyzed and transformed to 7-keto products. Optimal yields of 7-ketolithocholic acid and 7-ketodeoxycholic acid were obtained after 78 h of incubation. Culture pH changed with time and was characterized by an initial drop (1.1 pH units) and a gradual increase back to the starting pH (7.3). Corroborating these observations, an inducible, NADP-dependent, 7α-hydroxysteroid dehydrogenase was demonstrated in cell extracts of strain F-6. A trace of NAD-dependent 7α-hydroxysteroid dehydrogenase was also found. A substantial increase in the specific activity of the NADP-dependent 7α-hydroxysteroid dehydrogenase was observed when either 7-ketolithocholic acid, chenodeoxycholic acid, or deoxycholic acid was included in the growth medium. Optimal induction of the NADP-dependent 7α-hydroxysteroid dehydrogenase was achieved with 0.3–0.4 mM 7-ketolithocholic acid. Production of the enzyme(s) was optimal at 6–8 h of growth and the 7α-hydroxysteroid dehydrogenases had a pH optimum of approximately 11. The 7α-hydroxysteroid dehydrogenase from strain F-6 was purified 12-fold by triazine dye affinity chromatography with reactive blue 2 (Cibacron blue) agarose (95% yield). It was successfully lyophilized into a stable powder form.

scholarly journals Role of 11βHSD Type 2 Enzyme Activity in Essential Hypertension and Children with Chronic Kidney Disease (CKD)

2012 ◽  
Vol 97 (10) ◽  
pp. 3622-3629 ◽  
Author(s):  
Anil Mongia ◽  
Risa Vecker ◽  
Minu George ◽  
Anita Pandey ◽  
Hanan Tawadrous ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Blood Pressure ◽  
Essential Hypertension ◽  
Kidney Disease ◽  
Mineralocorticoid Receptor ◽  
Filtration Rate ◽  
Hydroxysteroid Dehydrogenase ◽  
Gas Chromatography Mass Spectrometry ◽  
Z Scores

Abstract Background: The mineralocorticoid receptor is protected from excess of glucocorticoids by conversion of active cortisol to inactive cortisone by enzyme 11β-hydroxysteroid dehydrogenase type 2 present in the kidney. The metabolites of cortisol and cortisone are excreted in the urine as tetrahydrocortisol (5αTHF+5βTHF) and tetrahydrocortisone (THE), respectively. Hypothesis: Patients with chronic kidney disease (CKD) and essential hypertension have a functional defect in their ability to convert cortisol to cortisone, thus leading to the activation of mineralocorticoid receptor. Objective: The objective of the investigation was to study the ratio of urinary steroids (5αTHF+5βTHF) to THE in patients with CKD, postrenal transplant, and essential hypertension and to compare the ratio with controls. Design/Methods: We enrolled 44 patients (17 with CKD, eight postrenal transplant, 19 with essential hypertension) and 12 controls. We measured spot urinary 5α-THF, 5β-THF, THE, free active cortisol and inactive cortisone by gas chromatography/mass spectrometry. We collected data on age, sex, cause of kidney disease, height, weight, body mass index, blood pressure, serum electrolytes, aldosterone, and plasma renin activity. Blood pressure percentiles and z-scores were calculated. The glomerular filtration rate was calculated using the modified Schwartz formula. Results: The ratios of 5αTHF+5βTHF to THE were significantly higher in patients with CKD [mean ± sd score (SDS) = 1.31 ± 1.07] as compared with essential hypertension (mean ± SDS = 0.59 ± 0.23; P = 0.02) and controls (mean ± SDS = 0.52 ± 0.25; P = 0.01). In the postrenal transplant group, the ratio was not significantly different (mean ± SDS = 0.71 ± 0.55). The urinary free cortisol to free cortisone ratios were significantly higher in the hypertension and CKD groups as compared with the controls. The 5αTHF+5βTHF to THE ratio negatively correlated with the glomerular filtration rate and positively correlated with systolic and diastolic blood pressure z-scores. The correlation of the blood pressure z-scores with ratios was stronger in the CKD group than the essential hypertension and posttransplant groups. Conclusions: We have elucidated a functional deficiency of 11β-hydroxysteroid dehydrogenase type 2 in children with CKD and a subset of essential hypertension. Urinary 5α-THF, 5β-THF, and THE analysis by gas chromatography/mass spectrometry should be a part of routine work-up of CKD and hypertensive patients.

Substrate and inhibitor specificity of the cloned human 11 beta-hydroxysteroid dehydrogenase type 2 isoform

1996 ◽  
Vol 270 (5) ◽  
pp. E900-E904 ◽  
Author(s):  
P. Ferrari ◽  
R. E. Smith ◽  
J. W. Funder ◽  
Z. S. Krozowski
Keyword(s):  
Dose Response ◽  
Mineralocorticoid Receptor ◽  
Response Curve ◽  
Hydroxysteroid Dehydrogenase ◽  
Significant Inhibition ◽  
Michaelis Constant ◽  
Intact Cells ◽  
Target Organs

The 11 beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2) enzyme is thought to confer specificity on the mineralocorticoid receptor by inactivating glucocorticoids in mineralocorticoid target organs. The cloned 11 beta HSD2 displayed Michaelis constant values for corticosterone and cortisol of 5.1 and 61 nM, respectively. Linearity in the dose-response curve ranged between 1 and 200 nM for corticosterone and 25 and 2,000 nM for cortisol, with no evidence for complex kinetics. Inhibition of cortisol oxidation by other steroids was purely competitive in nature. Inhibition of 11 beta HSD2 activity by the end product or aldosterone occurred only at supraphysiological levels, whereas corticosterone and deoxycorticosterone displayed significant inhibition at physiological concentrations and progesterone at concentrations that occur during pregnancy. In intact transfected CHOP cells, dexamethasone was converted to 11-dehydrodexamethasone by 11 beta HSD2 but not type 1 11 beta-hydroxysteroid dehydrogenase, an aspect that may be useful in evaluating 11 beta HSD activity in intact cells.

scholarly journals Differential Expression of Nuclear 11β-Hydroxysteroid Dehydrogenase Type 2 in Mineralocorticoid Receptor Positive and Negative Tissues

Endocrinology ◽  
1997 ◽  
Vol 138 (7) ◽  
pp. 3077-3080 ◽  
Author(s):  
Massimiliano D. Petrelli ◽  
Sylvia S. Lim-Tio ◽  
Jennifer Condon ◽  
Martin Hewison ◽  
Paul M. Stewart
Keyword(s):  
Differential Expression ◽  
Mineralocorticoid Receptor ◽  
Hydroxysteroid Dehydrogenase

scholarly journals Increased Expression of Mineralocorticoid Receptor and 11β-Hydroxysteroid Dehydrogenase Type 2 in Human Atria During Atrial Fibrillation

2010 ◽  
Vol 33 (1) ◽  
pp. 23-29 ◽  
Author(s):  
Pei De-An ◽  
Li Li ◽  
Xu Zhi-Yun ◽  
Huang Jin-Yu ◽  
Xu Zheng-Ming ◽  
...  
Keyword(s):  
Atrial Fibrillation ◽  
Mineralocorticoid Receptor ◽  
Hydroxysteroid Dehydrogenase

scholarly journals Two Homozygous Mutations in the 11β-Hydroxysteroid Dehydrogenase Type 2 Gene in a Case of Apparent Mineralocorticoid Excess

2003 ◽  
Vol 88 (6) ◽  
pp. 2501-2507 ◽  
Author(s):  
Cristian A. Carvajal ◽  
Alexis A. Gonzalez ◽  
Damian G. Romero ◽  
Angel González ◽  
Lorena M. Mosso ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Mineralocorticoid Receptor ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Hydroxysteroid Dehydrogenase ◽  
Protective Mechanism ◽  
Wild Type ◽  
Ovary Cells ◽  
Low Renin Hypertension

The human microsomal 11β-hydroxysteroid dehydrogenase type 2 (11βHSD2) metabolizes active cortisol into cortisone and protects the mineralocorticoid receptor from glucocorticoid occupancy. In a congenital deficiency of 11β-HSD2, the protective mechanism fails and cortisol gains inappropriate access to mineralocorticoid receptor, resulting in low-renin hypertension and hypokalemia. In the present study, we describe the clinical and molecular genetic characterization of a patient with a new mutation in the HSD11B2 gene. This is a 4-yr-old male with arterial hypertension. The plasma renin activity and serum aldosterone were undetectable in the presence of a high cortisol to cortisone ratio. PCR amplification and sequence analysis of HSD11B2 gene showed the homozygous mutation in exon 4 Asp223Asn (GAC→AAC) and a single nucleotide substitution C→T in intron 3. Using site-directed mutagenesis, we generated a mutant 11βHSD2 cDNA containing the Asp223Asn mutation. Wild-type and mutant cDNA was transfected into Chinese hamster ovary cells and enzymatic activities were measured using radiolabeled cortisol and thin-layer chromatography. The mRNA and 11βHSD2 protein were detected by RT-PCR and Western blot, respectively. Wild-type and mutant 11βHSD2 protein was expressed in Chinese hamster ovary cells, but the mutant enzyme had only 6% of wild-type activity. In silico 3D modeling showed that Asp223Asn changed the enzyme’s surface electrostatic potential affecting the cofactor and substrate enzyme-binding capacity. The single substitution C→T in intron 3 (IVS3 + 14 C→T) have been previously reported that alters the normal splicing of pre-mRNA, given a nonfunctional protein. These findings may determine the full inactivation of this enzyme, explaining the biochemical profile and the early onset of hypertension seen in this patient.

Sign in / Sign up

Export Citation Format

Share Document