Showing papers on "Hydroxysteroid dehydrogenase published in 2004"
TL;DR: The characterization of a high-titer isoform specific monoclonal antibody (Ab) for AKR1C3 is described, which can be used to detect AKR 1C3 expression by immunohistochemistry in sections of paraffin-embedded mammary gland and prostate.
122 citations
TL;DR: The data support the possibility that alterations in splanchnic cortisol production contribute to visceral fat accumulation and the hepatic insulin resistance of obesity or type 2 diabetes.
Abstract: Glucocorticoids are potent regulators of protein, fat, and carbohydrate metabolism. To determine if cortisol production occurs within the splanchnic bed in humans, 11 nondiabetic subjects were studied using the hepatic/leg catheterization method along with an infusion of [9,11,12,12- 2 H 4 ] cortisol (D4-cortisol) as proposed by Andrews et al. In the fasting state, there was net release ( P P P P P P P 2 H 3 ] cortisone (D3-cortisone), which in turn generates [9,12,12 2 H 3 ] cortisol (D3-cortisol) via 11-β hydroxysteroid dehydrogenase (11β-HSD) type 1, D3-cortisol production can be used as an index of 11β-HSD type 1 activity. Net splanchnic D3-cortisol release (3.9 ± 0.4 μg/min) and splanchnic D3-cortisol production (7.1 ± 0.7 μg/min) occurred ( P r = 0.84; P r = 0.97; P
109 citations
TL;DR: Human adipose tissue is capable of active androgen synthesis catalysed by 17beta-HSD5, and increased expression in obesity may contribute to circulating androgen excess.
Abstract: Women with polycystic ovary syndrome (PCOS) have high circulating androgens, thought to originate from ovaries and adrenals, and frequently suffer from the metabolic syndrome including obesity. However, serum androgens are positively associated with body mass index (BMI) not only in PCOS, but also in simple obesity, suggesting androgen synthesis within adipose tissue. Thus we investigated androgen generation in human adipose tissue, including expression of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) isozymes, important regulators of sex steroid metabolism. Paired omental and subcutaneous fat biopsies were obtained from 27 healthy women undergoing elective abdominal surgery (age range 30-50 years; BMI 19.7-39.2 kg/m(2)). Enzymatic activity assays in preadipocyte proliferation cultures revealed effcient conversion of androstenedione to testosterone in both subcutaneous and omental fat. RT-PCR of whole fat and preadipocytes of subcutaneous and omental origin showed expression of 17beta-HSD types 4 and 5, but no relevant expression of 17beta-HSD types 1, 2, or 3. Microarray analysis confirmed this expression pattern (17beta-HSD5>17beta-HSD4) and suggested a higher expression of 17beta-HSD5 in subcutaneous fat. Accordingly, quantitative real-time RT-PCR showed significantly higher expression of 17beta-HSD5 in subcutaneous compared with omental fat (P<0.05). 17beta-HSD5 expression in subcutaneous, but not omental, whole fat correlated significantly with BMI (r=0.51, P<0.05). In keeping with these findings, 17beta-HSD5 expression in subcutaneous fat biopsies from six women taking part in a weight loss study decreased significantly with weight loss (P<0.05). A role for 17beta-HSD5 in adipocyte differentiation was further supported by the observed increase in 17beta-HSD5 expression upon differentiation of stromal preadipocytes to mature adipocytes (n=5; P<0.005), which again was higher in cells of subcutaneous origin. Functional activity of 17beta-HSD5 also significantly increased with differentiation, revealing a net gain in androgen activation (androstenedione to testosterone) in subcutaneous cultures, contrasting with a net gain in androgen inactivation (testosterone to androstenedione) in omental cultures. Thus, human adipose tissue is capable of active androgen synthesis catalysed by 17beta-HSD5, and increased expression in obesity may contribute to circulating androgen excess.
74 citations
TL;DR: In the human prostate, UGT 2B15 and UGT2B17 genes have complementary roles, and are expressed in cells where their specific substrates are synthesized, which reinforces the hypothesis that UGT enzymes catalyze an important mechanism for modulating the action of steroids and protecting the steroid‐sensitive tissues from deleteriously high steroid concentrations.
Abstract: In humans, 3β‐hydroxysteroid dehydrogenase (3β‐HSD), 17β‐HSD, and 5α‐reductase enzymes convert dehydroepiandrosterone (DHEA), androstenedione, and testosterone into the most potent natural androgen dihydrotestosterone (DHT) in the prostate. This androgen is transformed mainly in situ to two Phase I metabolites, androsterone (ADT) and androstane‐3α,17β‐diol (3α‐DIOL), which can, however, be back‐converted to DHT. Here, we report recent findings on the characterization of specific anti‐UDP‐glucuronosyltransferases (UGT)2B15 and 2B17 antibodies and their use to identify UGT2B expressing‐cells in the human prostate epithelium. We found that UGT2B17 is expressed in basal cells where DHEA is converted into 3α‐DIOL and ADT. By contrast, the expression of UGT2B15 was observed only in luminal cells, where DHT is formed from testosterone. These results demonstrate that, in the human prostate, UGT2B15 and UGT2B17 genes have complementary roles, and are expressed in cells where their specific substrates are synthesiz...
61 citations
TL;DR: Unexpectedly, a search of the Takifugu, Tetraodon and medaka genomes only found an ortholog to 11β‐HSD2 and none to 17β‐ HSD2, while the zebrafish genome contains orthologs of both enzymes.
55 citations
TL;DR: It is concluded that 17βHSD types 1, 2, and 3 catalyze both oxidative and reductive reactions in HEK-293 cells at intrinsic rates that are much faster than those estimated from single-isotope studies.
54 citations
TL;DR: Combining structural, site-directed mutagenesis, kinetic and fluorescence titration studies, it is found that the selectivity of rb20alpha-HSD is mediated by the relaxation of loop B (residues 223-230), partly controlled by the nature of residue 230.
39 citations
Patent•
21 May 2004
TL;DR: In this article, a method for promoting wound healing was proposed, which consisted of administering to a mammal, including man, in need of such promotion an effective amount of an inhibitor of 11-s-hydroxysteroid dehydrogenase type 1.
Abstract: The invention relates to a method for promoting wound healing, said method comprising administering to a mammal, including man, in need of such promotion an effective amount of an inhibitor of 11-s-hydroxysteroid dehydrogenase type 1 wherein the said l ls-HSD1 inhibitor has the formula (I) wherein T, R1, A1 and A2 are as defined in the specification. These compounds may also be used in the manufacture of a medicament for promoting wound healing.
32 citations
TL;DR: The present data indicate that E2 can be formed through the action of 17beta-HSD type 1 in specific cells of the gonads and peripheral tissues, and is probably limited to the cells involved in its formation in an intracrine fashion.
Abstract: The enzyme 17-hydroxysteroid dehydrogenase (17-HSD) type 1 catalyzes the conversion of estrone (E1) into 17 estradiol (E2). To gain information about the cellular localization of 17-HSD mRNA type 1 expression, we performed in situ hybridization using a 35 S-labeled cRNA probe in several tissues of adult mice of both sexes. In the ovary, high expression was found in granulosa cells of growing follicles. No specific labeling could be observed in corpora lutea or interstitial cells. In the pituitary gland of animals of both sexes, 17-HSD type 1 mRNA was expressed in the intermediate lobe melanotrophs while no specific signal could be detected in the anterior or posterior lobes of the pituitary. In the prostate, 17-HSD type 1 mRNA was exclusively found in the epithelial cells. In both male and female mouse dorsal skin, a specific hybridization signal was seen in the sebaceous glands while the epidermis, stroma, hair follicles and sweat glands were unlabeled. In the testis, a hybridization signal was detected in germ cells of the seminiferous tubules, Leydig cells being unlabeled. The present data indicate that E2 can be formed through the action of 17-HSD type 1 in specific cells of the gonads and peripheral tissues. In the testes and peripheral tissues, the action of E2 is probably limited to the cells involved in its formation in an intracrine fashion.
29 citations
TL;DR: Partial 11-&bgr; hydroxysteroid dehydrogenase type 2 deficiency is pointed to as a novel feature of essential hypertension, which should stimulate search for new signaling pathways and therapeutical targets.
Abstract: The enzyme 11-beta hydroxysteroid dehydrogenase type 2 plays a major role in blood pressure regulation. It metabolizes glucocorticoid hormones into derivatives with low affinity for the mineralocorticoid receptor, preventing its permanent occupancy by circulating cortisol, which is 100- to 1000-fold more abundant than aldosterone in the plasma. Inactivating mutations of the enzyme result in severe hypertension, as seen in children with apparent mineralocorticoid excess syndrome. In patients with essential hypertension, however, attempts to evidence enzyme deficiency have been inconclusive. In this pilot study, its catalytic activity was measured directly in aldosterone-sensitive sweat gland ducts collected from skin biopsy samples of 10 male normotensive subjects and 10 subjects with essential hypertension (more than 140 to 90 mm Hg) with no sign of hypermineralocorticism. Isolated ducts were assayed for nicotinamide-dinucleotide-dependent dehydrogenase activity (transformation of tritiated corticosterone into tritiated-11 dehydrocorticosterone, as measured by high-pressure liquid chromatography). Hypertensive patients exhibited significantly lower 11-beta hydroxysteroid dehydrogenase type 2 activity (9.7+/-4.7 femtomoles per 3 mm length of duct and per 10 minutes incubation, median+/-SD) than did normotensive subjects (15.9+/-2.6). Such defect was undetectable using the classical urinary corticosteroid metabolism indexes, probably because of compensatory mechanisms. Relations between these findings and blood pressure levels should benefit from direct enzyme measurements in the vasculature. In conclusion, this cross-sectional study points to partial 11-beta hydroxysteroid dehydrogenase type 2 deficiency as a novel feature of essential hypertension, which should stimulate search for new signaling pathways and therapeutical targets.
29 citations
TL;DR: Findings suggest new roles for both peroxisome proliferator-activated receptor α and 17β-hydroxysterol dehydrogenase type 11 in lipid metabolism and/or detoxification in the intestine.
Abstract: In order to study the role of peroxisome proliferator-activated receptor α in mouse intestine, its agonist-induced proteins were identified by peptide mass fingerprinting followed by Northern blot analysis using their cDNAs One of the most remarkably induced proteins was identified as 17β-hydroxysterol dehydrogenase type 11 Its very rapid induction by various agonists was most efficient in intestine and then in liver These findings together with recently reported results showing the enzyme family's wide substrate spectrum, including not only glucocorticoids and sex steroids but also bile acids, fatty acids and branched chain amino acids, suggest new roles for both peroxisome proliferator-activated receptor α and 17β-hydroxysterol dehydrogenase type 11 in lipid metabolism and/or detoxification in the intestine
TL;DR: In a series of cinnamates and related coumarin-3-carboxylates, a number of compounds proved to be potent inhibitors of both the oxidative and reductive reactions catalyzed by 17beta-HSDcl, with IC(50) values in the low micromolar range.
TL;DR: Results demonstrate that the gonadotropin-dependent induction of follicular luteinization is accompanied by an increase in 17betaHSD4 expression, which appears as a potential complementary mechanism to reduce circulating levels of 17beta-estradiol after the LH surge.
Abstract: 17β-Hydroxysteroid dehydrogenase type 4 (17βHSD4) has a unique multidomain structure, with one domain involved in 17β-estradiol inactivation. The objective of the study was to investigate the regulation of 17βHSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17βHSD4 cDNA was cloned and was shown to encode a 735-amino acid protein that is highly conserved (81–87% identity) compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17βHSD4 transcripts in equine preovulatory follicles isolated between 0–39 h after hCG treatment. Results showed the presence of basal 17βHSD4 mRNA expression before hCG treatment, but an increase was observed in follicles obtained 24 h after hCG (P < 0.05). Analyses of isolated preparations of granulosa and theca interna cells identified basal mRNA expression in both layers, but granulosa cells appeared as the predominant site of follicular 17βHSD4 mRNA induction. A specific polyclonal...
TL;DR: The present study suggests that HSDs may catalyze the C-3 epimerization of vitamin D compounds and modulate their concentrations and biological activities in animals and humans.
TL;DR: The substantially higher affinities of human 3 β‐HSD1 for substrate and inhibitor steroids measured using the pure isoenzymes have been validated using microsome‐bound 3β‐ HSD1 and 3β-HSD2 in the MCF‐7 cells.
Abstract: Two distinct genes encode the tissue-specific expression of the two isoforms of human 3beta-hydroxysteroid dehydrogenase: 3beta-HSD1 (placenta, mammary gland, breast tumors) and 3beta-HSD2 (gonads, adrenals). Purified 3beta-HSD1 utilizes DHEA as a substrate with 13-fold lower Km than 3beta-HSD2. Using homogenates of human MCF-7 Tet-off breast tumor cells stably transfected with human 3beta-HSD1 or 3beta-HSD2, DHEA is utilized as substrate by 3beta-HSD1 (Km = 4.8 microM) much more avidly than by 3beta-HSD2 (Km = 43 microM). In addition, the 3beta-HSD inhibitor, epostane, binds to purified 3beta-HSD1 with a 17-fold higher affinity compared to 3beta-HSD2. In the MCF-7 cells, two 3beta-HSD inhibitors block 3beta-HSD1 activity (Ki = 0.06 microM, epostane; 0.08 microM, trilostane) with 12- to 14-fold higher affinities compared to the inhibition of 3beta-HSD2 (Ki = 0.85 microM, epostane; 1.01 microM, trilostane). Thus, the substantially higher affinities of human 3beta-HSD1 for substrate and inhibitor steroids measured using the pure isoenzymes have been validated using microsome-bound 3beta-HSD1 and 3beta-HSD2 in the MCF-7 cells. Similar to our previously reported H156Y mutant of 3beta-HSD1, the Q105M mutant of 3beta-HSD1 shifts the substrate and inhibitor kinetic profiles to those of wild-type 3beta-HSD2. However, the Q105M mutant of 3beta-HSD2 retains the substrate and inhibitor kinetic profiles of wild-type 3beta-HSD2. Our structural homology model of human 3beta-HSD predicts that Gln105 on one enzyme subunit hydrogen-binds to His156 on the other subunit of the enzyme homodimer. The higher affinity of 3beta-HSD1 for the steroids may be related to different subunit interactions in the quaternary structures of the two isoenzymes. It may be possible to exploit these kinetic differences to selectively inhibit the conversion of DHEA ultimately to estradiol by 3beta-HSD1 and slow the growth of breast tumor cells.
TL;DR: The results suggest that progesterone can be inactivated by 20alpha-HSD in some specific brain areas.
TL;DR: It is demonstrated that the mouse 3β–HSD I and VI isoforms can inactivate testosterone within an intact cell milieu, important not only in establishment of structure‐function relationships, but also whenever murine systems are used for developmental/reproductive paradigms associated with human disorders.
Abstract: 3β–Hydroxysteroid dehydrogenase (3β‐HSD) activity is essential for the synthesis of all classes of steroid hormones, converting various Δ5–3β–hydroxysteroids into hormonally active Δ4–3–ketosteroids in NAD+–dependent reactions. Certain 3β–HSD isoforms have been reported to exhibit additional dehydrogenase character (e.g., 17–hydroxysteroid dehydrogenase/reductase). We have investigated whether mouse type I (adrenal/gonadal) and type VI 3β–HSDs (uterine/embryonic) display significant 17β–HSD–like activity. Nonsteroidogenic HEK 293T cells were transiently transfected with pCMV–based expression vectors containing mouse type I and type VI 3β–HSDs. Transfected cells expressing either mouse type I or type VI 3β–HSD converted testosterone to androstenedione, albeit at rates one‐tenth of those of pregnenolone to progesterone in similarly transfected 293T cells. Our findings demonstrate that the mouse 3β–HSD I and VI isoforms can inactivate testosterone within an intact cell milieu. These findings are important no...
TL;DR: Substrate specificity demonstrated that 3-Hydroxyhexobarbital dehydrogenases dehydrogenate not only α,β-unsaturated cyclic and acyclic secondary alcohols but also some 17β-, 3α-hydroxysteroids or both, except for the human enzyme that had a molecular weight of about 58000 and used NAD+ alone.
Abstract: Hexobarbital, a short-acting hypnotic, is metabolized to 3′-hydroxyhexobarbital by cytochrome P450, and then to 3′-oxohexobarbital by liver cytosolic dehydrogenase. New methods of separation for hexobarbital and its metabolites by TLC have been developed and applied to study the metabolism of hexobarbital enantiomers and stereoselective metabolism of hexobarbital. (+)-Hexobarbital preferentially was transformed into β-3′-hydroxyhexobarbital and the (−)-enantiomer preferentially transformed into α-3′-hydroxyhexobarbital by rat liver microsomes. Glucuronidation and dehydrogenation of 3′-hydroxyhexobarbital were also stereoselective and the S-configuration at the 3′-position was preferred. α-3′-Hydroxyhexobarbital from (−)-hexobarbital and the β-isomer from (+)-hexobarbital were shown to be preferentially conjugated with glucuronic acid in rabbit urine, and to be preferentially dehydrogenated to form 3′-oxohexobarbital by rabbit and guinea pig 3-hydroxyhexobarbital dehydrogenases. A new metabolic pathway of hexobarbital was found in which 3′-oxohexobarbital reacts with glutathione to form 1,5-dimethylbarbituric acid and a cyclohexenone-glutathione adduct, a novel metabolite. 1,5-Dimethylbarbituric acid was excreted into the urine and the cyclohexenone-glutathione adduct into the bile of rats dosed with hexobarbital. 3-Hydroxyhexobarbital dehydrogenases that dehydrogenate 3-hydroxyhexobarbital into 3′-oxohexobarbital were purified from the liver cytosol of rabbits, guinea pigs, goats, rats, mice, hamsters, and humans and characterized. These enzymes were monomeric proteins and had molecular weights of about 34500—42000, and used NAD+ and NADP+ as cofactors, except for the human enzyme that had a molecular weight of about 58000 and used NAD+ alone. Each enzyme exhibited its own characteristics. Substrate specificity demonstrated that 3-hydroxyhexobarbital dehydrogenases dehydrogenate not only α,β-unsaturated cyclic and acyclic secondary alcohols but also some 17β-, 3α-hydroxysteroids or both, except for the human enzyme. The amino acid sequence of the hamster enzyme indicated that it belongs to the aldo-keto reductase superfamily and hydroxysteroid dehydrogenase subfamily.
TL;DR: The amino acid sequence of the N-terminal of the purified enzyme showed that 7β- HSD had sequence similarity to rabbit type I 11β-hydroxysteroid dehydrogenase (11β-HSD), indicating that 7 β-HSS may belong to the rabbit typeI 11β -HSD family and may play the same role in the metabolism of 11-Hydroxysteroids and 7-hydroxsterols.
01 Mar 2004
TL;DR: A novel 7α-hydroxysteroid dehydrogenase (7α-HSD) was purified from Pseudomonas sp.
Abstract: A novel 7α-hydroxysteroid dehydrogenase (7α-HSD) was purified from Pseudomonas sp. B-0831. The molecular weight of the purified enzyme was 25 k on SDS-PAGE and 108 k on gel filtration analysis, suggesting that the enzyme exists as a tetramer of an identical subunit, similar to those of 7α-HSD from E. coli HB101 and Eubacterium sp. VPI 12708. 7α-HSD from P. sp. B-0831 showed high NAD + -dependence and was able to catalyze the oxido-reduction of 7α-hydroxy bile acids including glycine and taurine conjugates. The Km value for cholic acid was determined to be 0.25 mM, which is similar to that for chenodeoxycholic acid and is about four times smaller than those for the conjugates of cholic acid. The kcat values for the conjugates were determined to be about 60-70% of that for free cholic acid. In addition to NAD + , 7α-HSD from P. sp. B-0831 can utilize thio-NAD + to the same extent.
TL;DR: In this article, a new family of nonsteroidal inhibitors of 17β-hydroxysteroid dehydrogenase, designed from flavones and chalcones, was presented.
Abstract: We present the synthesis of a new family of nonsteroidal inhibitors of 17beta-hydroxysteroid dehydrogenase, designed from flavones and chalcones. Their inhibitory potential was screened on 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17beta-HSDcl), a model enzyme of the short-chain dehydrogenase/reductase superfamily. In a series of cinnamates and related coumarin-3-carboxylates, a number of compounds proved to be potent inhibitors of both the oxidative and reductive reactions catalyzed by 17beta-HSDcl, with IC(50) values in the low micromolar range.
Patent•
13 Dec 2004TL;DR: In this paper, substituted thiophenepyrimidinone derivatives and their use in therapy, especially for use in the treatment and/or prevention of a steroid hormone dependent diseases or disorder, such as steroid hormone-dependent diseases or disorders requiring inhibition of 17β-hydroxysteroid dehydrogenase enzymes.
Abstract: This invention relates to novel substituted thiophenepyrimidinone derivatives and their use in therapy, especially for use in the treatment and/or prevention of a steroid hormone dependent diseases or disorder, such as steroid hormone dependent diseases or disorders requiring inhibition of 17β-hydroxysteroid dehydrogenase enzymes.
Patent•
07 Jun 2004TL;DR: Thiopheneprymidinone compounds are useful in therapy, especially for use in the treatment and/or prevention of a steroid hormone dependent disorder, or disorder requiring the inhibition of a 17β-hydroxysteroid dehydrogenase (17β-HSD).
Abstract: Thiopheneprymidinone compounds useful in therapy, especially for use in the treatment and/or prevention of a steroid hormone dependent disorder, preferably a steroid hormone dependent disease or disorder requiring the inhibition of a 17β-hydroxysteroid dehydrogenase (17β-HSD) such as 17β-HSD type 1, type 2 or type 3 enzyme.
01 Nov 2004
Patent•
09 Nov 2004
TL;DR: In this article, the authors describe 3.15-substituted estrone compounds which act as inhibitors of 17β-hydroxysteroid dehydrogenase type I (17β-HSD1) enzymes.
Abstract: 3,15-substituted estrone compounds which act as inhibitors of 17β-hydroxysteroid dehydrogenase type I (17β-HSD1), salts thereof, pharmaceutical preparations containing such compounds, processes for preparing such compounds, and therapeutic uses of such compounds, particularly in the treatment or inhibition of steroid hormone dependent diseases or disorders, such as steroid hormone dependent diseases or disorders requiring the inhibition of 17β-hydroxysteroid dehydrogenase type I enzymes and/or requiring the lowering of the endogenous 17β-estradiol concentration, as well as the general use of selective 17β-hydroxysteroid dehydrogenase type 1 inhibitors which possess in addition no or only pure antagonistic binding affinities to the estrogen receptor for the treatment or inhibition of benign gynecological disorders, particularly endometriosis.
Patent•
16 Dec 2004
TL;DR: In this article, novel compositions useful for the treatment of androgen dependant diseases, comprising hydroxysteroid dehydrogenase inhibitors adsorbed on silica or microcrystalline cellulose, were presented.
Abstract: Disclosed herein are novel compositions useful for the treatment of androgen dependant diseases, comprising hydroxysteroid dehydrogenase inhibitors adsorbed on silica or microcrystalline cellulose.
Patent•
27 May 2004TL;DR: In this article, the authors used 3-beta-hydroxysteroid dehydrogenase-isomerase gene products to prevent, ameliorate or correct dysfunctions or diseases including, but not limited to, cancer.
Abstract: Reagents that regulate human 3-beta-hydroxysteroid dehydrogenase-isomerase and reagents which bind to human 3-beta-hydroxysteroid dehydrogenase-isomerase gene products can play a role in preventing, ameliorating, or correcting dysfunctions or diseases including, but not limited to, cancer.