scholarly journals Structural and functional comparison of two human liver dihydrodiol dehydrogenases associated with 3 α-hydroxysteroid dehydrogenase activity

1992 ◽  
Vol 282 (3) ◽  
pp. 741-746 ◽  
Author(s):  
Y Deyashiki ◽  
H Taniguchi ◽  
T Amano ◽  
T Nakayama ◽  
A Hara ◽  
...  
Keyword(s):  
Bile Acids ◽  
Dehydrogenase Activity ◽  
Human Liver ◽  
Hydrogen Transfer ◽  
Peptide Mapping ◽  
Reductase Activity ◽  
High Specificity ◽  
Hydroxysteroid Dehydrogenase ◽  
Inflammatory Drugs ◽  
Anti Inflammatory

Two monomeric dihydrodiol dehydrogenases with pI values of 5.4 and 7.6 were co-purified with androsterone dehydrogenase activity to homogeneity from human liver. The two enzymes differed from each other on peptide mapping and in their heat-stabilities; with respect to the latter the dihydrodiol dehydrogenase and 3 alpha-hydroxysteroid dehydrogenase activities of the respective enzymes were similarly inactivated. The pI 5.4 enzyme was equally active towards trans- and cis-benzene dihydrodiols, and towards (S)- and (R)-forms of indan-1-ol and 1,2,3,4-tetrahydronaphth-1-ol and oxidized the 3 alpha-hydroxy group of C19-, C21- and C24-steroids, whereas the pI 7.6 enzyme showed high specificity for trans-benzene dihydrodiol, (S)-forms of the alicyclic alcohols and C19- and C21-steroids. Although the two enzymes reduced various xenobiotic carbonyl compounds and the 3-oxo group of C19- and C21-steroids, and were A-specific in the hydrogen transfer from NADPH, only the pI 5.4 enzyme showed reductase activity towards 7 alpha-hydroxy-5 beta-cholestan-3-one and dehydrolithocholic acid. The affinity of the two enzymes for the steroidal substrates was higher than that for the xenobiotic substrates. The two enzymes also showed different susceptibilities to the inhibition by anti-inflammatory drugs and bile acids. Whereas the pI-5.4 enzyme was highly sensitive to anti-inflammatory steroids, showing mixed-type inhibitions with respect to indan-1-ol and androsterone, the pI 7.6 enzyme was inhibited more potently by non-steroidal anti-inflammatory drugs and bile acids than by the steroidal drugs, and the inhibitions were all competitive. These structural and functional differences suggest that the two enzymes are 3 alpha-hydroxysteroid dehydrogenase isoenzymes.

GLUCURONIDATION OF NONSTEROIDAL ANTI-INFLAMMATORY DRUGS: IDENTIFYING THE ENZYMES RESPONSIBLE IN HUMAN LIVER MICROSOMES

2005 ◽  
Vol 33 (7) ◽  
pp. 1027-1035 ◽  
Author(s):  
Gwendolyn E. Kuehl ◽  
Johanna W. Lampe ◽  
John D. Potter ◽  
Jeannette Bigler
Keyword(s):  
Human Liver ◽  
Liver Microsomes ◽  
Human Liver Microsomes ◽  
Inflammatory Drugs ◽  
Anti Inflammatory

'Liver-type' 11β-hydroxysteroid dehydrogenase cDNA encodes reductase but not dehydrogenase activity in intact mammalian COS-7 cells

1994 ◽  
Vol 13 (2) ◽  
pp. 167-174 ◽  
Author(s):  
S C Low ◽  
K E Chapman ◽  
C R W Edwards ◽  
J R Seckl
Keyword(s):  
Dehydrogenase Activity ◽  
Rat Liver ◽  
Mammalian Cells ◽  
Reductase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Luciferase Reporter ◽  
Luciferase Expression ◽  
Intact Cells ◽  
Mmtv Ltr

ABSTRACT 11β-Hydroxysteroid dehydrogenase (11β-HSD) catalyses the metabolism of corticosterone to inert 11-dehydrocorticosterone, thus preventing glucocorticoid access to otherwise non-selective renal mineralocorticoid receptors (MRs), producing aldosterone selectivity in vivo. At least two isoforms of 11β-HSD exist. One isoform (11β-HSD1) has been purified from rat liver and an encoding cDNA cloned from a rat liver library. Transfection of rat 11β-HSD1 cDNA into amphibian cells with a mineralocorticoid phenotype encodes 11 β-reductase activity (activation of inert 11-dehydrocorticosterone) suggesting that 11β-HSD1 does not have the necessary properties to protect renal MRs from exposure to glucocorticoids. This function is likely to reside in a second 11β-HSD isoform. 11β-HSD1 is co-localized with glucocorticoid receptors (GRs) and may modulate glucocorticoid access to this receptor type. To examine the predominant direction of 11β-HSD1 activity in intact mammalian cells, and the possible role of 11β-HSD in regulating glucocorticoid access to GRs, we transfected rat 11β-HSD1 cDNA into a mammalian kidney-derived cell system (COS-7) which has little endogenous 11β-HSD activity or mRNA expression. Homogenates of COS-7 cells transfected with increasing amounts of 11β-HSD cDNA exhibited a dose-related increase in 11 β-dehydrogenase activity. In contrast, intact cells did not convert corticosterone to 11-dehydrocorticosterone over 24 h, but showed a clear dose-related 11β-reductase activity, apparent within 4 h of addition of 11-dehydrocorticosterone to the medium. To demonstrate that this reflected a change in functional intracellular glucocorticoids, COS-7 cells were co-transfected with an expression vector encoding GR and a glucocorticoid-inducible MMTV-LTR luciferase reporter construct, with or without 11β-HSD. Corticosterone induced MMTV-LTR luciferase expression in the presence or absence of 11β-HSD. 11-Dehydrocorticosterone was without activity in the absence of 11β-HSD, but induced MMTV-LTR luciferase activity in the presence of 11β-HSD. These results indicate that rat 11β-HSD1 can behave exclusively as a reductase in intact mammalian cells. Thus in some tissues in vivo, 11β-HSD1 may regulate ligand access to GRs by reactivating inert glucocorticoids.

scholarly journals Human liver class I alcohol dehydrogenaseγγ isozyme: the sole cytosolic 3β-hydroxysteroid dehydrogenase of iso bile acids

Hepatology ◽  
2000 ◽  
Vol 31 (4) ◽  
pp. 990-996 ◽  
Author(s):  
Hanns-Ulrich Marschall ◽  
Udo C.I. Oppermann ◽  
Stefan Svensson ◽  
Erik Nordling ◽  
Bengt Persson ◽  
...  
Keyword(s):  
Bile Acids ◽  
Human Liver ◽  
Hydroxysteroid Dehydrogenase ◽  
Class I

EFFECTS OF TAMOXIFEN ON THE BINDING AND METABOLISM OF TESTOSTERONE BY HUMAN PROSTATIC TISSUE AND PLASMA IN VITRO

1979 ◽  
Vol 83 (3) ◽  
pp. 369-378 ◽  
Author(s):  
F. K. HABIB ◽  
G. RAFATI ◽  
M. R. G. ROBINSON ◽  
S. R. STITCH
Keyword(s):  
Dehydrogenase Activity ◽  
Reductase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Inhibitory Effect ◽  
Prostatic Tissue ◽  
Sex Hormone Binding Globulin ◽  
Benign Tissue ◽  
The Mean ◽  
Malignant Prostate

The in-vitro metabolism of testosterone in benign and malignant prostatic tissue was examined and distinct quantitative differences between the two types of specimens were observed. The major metabolite of testosterone in the hyperplastic prostate was 5α-dihydrotestosterone and a high 3α(β)-hydroxysteroid dehydrogenase activity was also detected. In the malignant tissue, 5α-reductase activity was considerably reduced and there was little or no androstanediol formed; the 17β-dehydrogenase activity was, however, higher than in the benign tissue. The decrease in 5α-reductase was always followed by a compensatory change in the 3α(β)-hydroxysteroid dehydrogenase of the malignant prostate. The present study revealed that the ratio of the mean activities of 5α-reductase to 3α(β)-hydroxysteroid dehydrogenase in the two types of specimen always remained a constant. Although the antioestrogen, tamoxifen, induced an inhibitory effect on the activities of 5α-reductase and 17β-hydroxysteroid dehydrogenase in the gland, the present investigation also suggested that tamoxifen stimulated the activity of 3α(3β)-hydroxysteroid dehydrogenase. In blood, the action of tamoxifen appeared to be confined to the displacement of androgens from the binding sites on the sex hormone binding globulin.

scholarly journals Effect of indomethacin on bile acid-phospholipid interactions: implication for small intestinal injury induced by nonsteroidal anti-inflammatory drugs

2010 ◽  
Vol 298 (5) ◽  
pp. G722-G731 ◽  
Author(s):  
Yong Zhou ◽  
Elizabeth J. Dial ◽  
Rand Doyen ◽  
Lenard M. Lichtenberger
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Membrane Surface ◽  
Intestinal Injury ◽  
Disruptive Effect ◽  
Plasma Membrane Permeability ◽  
Small Intestinal Injury ◽  
Inflammatory Drugs ◽  
Small Intestinal ◽  
Anti Inflammatory

The injurious effect of nonsteroidal anti-inflammatory drugs (NSAIDs) in the small intestine was not appreciated until the widespread use of capsule endoscopy. Animal studies found that NSAID-induced small intestinal injury depends on the ability of these drugs to be secreted into the bile. Because the individual toxicity of amphiphilic bile acids and NSAIDs directly correlates with their interactions with phospholipid membranes, we propose that the presence of both NSAIDs and bile acids alters their individual physicochemical properties and enhances the disruptive effect on cell membranes and overall cytotoxicity. We utilized in vitro gastric AGS and intestinal IEC-6 cells and found that combinations of bile acid, deoxycholic acid (DC), taurodeoxycholic acid, glycodeoxycholic acid, and the NSAID indomethacin (Indo) significantly increased cell plasma membrane permeability and became more cytotoxic than these agents alone. We confirmed this finding by measuring liposome permeability and intramembrane packing in synthetic model membranes exposed to DC, Indo, or combinations of both agents. By measuring physicochemical parameters, such as fluorescence resonance energy transfer and membrane surface charge, we found that Indo associated with phosphatidylcholine and promoted the molecular aggregation of DC and potential formation of larger and isolated bile acid complexes within either biomembranes or bile acid-lipid mixed micelles, which leads to membrane disruption. In this study, we demonstrated increased cytotoxicity of combinations of bile acid and NSAID and provided a molecular mechanism for the observed toxicity. This mechanism potentially contributes to the NSAID-induced injury in the small bowel.

scholarly journals Dual Effects of Anti-inflammatory 2-Arylpropionic Acid Derivatives on a Major Isoform of Human Liver 3.ALPHA.-Hydroxysteroid Dehydrogenase.

1998 ◽  
Vol 21 (11) ◽  
pp. 1148-1153 ◽  
Author(s):  
Tomohiro YAMAMOTO ◽  
Kazuya MATSUURA ◽  
Shunichi SHINTANI ◽  
Akira HARA ◽  
Yoshiyuki MIYABE ◽  
...  
Keyword(s):  
Human Liver ◽  
Hydroxysteroid Dehydrogenase ◽  
Arylpropionic Acid ◽  
Anti Inflammatory
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