Biomedical subjects

M Axelson

Publications and source records attributed to M Axelson.

At least 37 records · Page 2Linked to original sources

PubMed1996

27-hydroxylated low density lipoprotein (LDL) cholesterol can be converted to 7alpha,27-dihydroxy-4-cholesten-3-one (cytosterone) before suppressing cholesterol production in normal human fibroblasts. Evidence that an altered metabolism of ldl cholesterol can underlie a defective feedback control in malignant cells.

The formation of oxysterols in cultured human fibroblasts and their physiological roles as intracellular regulators of cholesterol production have been investigated. In the presence of low density lipoproteins (LDL), normal fibroblasts converted LDL cholesterol to 27hydroxycholesterol, which was further metabolized to 7alpha, 27-dihydroxycholesterol, 7alpha,27-dihydroxy-4-cholesten-3-one, and 7alpha-hydroxy-3-oxo-4-cholestenoic acid. Autooxidation products of cholesterol contaminating the lipoproteins were also metabolized in the cells. 7alpha-Hydroxycholesterol was converted to 7alpha-hydroxy-4-cholesten-3-one prior to 27-hydroxylation and further oxidation to 7alpha-hydroxy-3-oxo-4-cholestenoic acid. 7beta-Hydroxycholesterol and 7-oxocholesterol were 27-hydroxylated and then oxidized to C27-acids. Oxidation of the 7beta-hydroxy group also occurred. 25-Hydroxycholesterol was 7alpha-hydroxylated and further oxidized to 7alpha,25-dihydroxy-4-cholesten-3-one. 25-Hydroxylation of sterols was observed only under specific conditions. In contrast, only small amounts of oxysterols were formed in virus-transformed human fibroblasts when incubated with lipoproteins. This was due to very low activities of the 27- and 7alpha-hydroxylating enzymes. The rate of oxidation at C-3 was also decreased moderately. A defective suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase by LDL and autooxidation products of cholesterol observed in the transformed fibroblasts could be caused by the deficiencies of the sterol-metabolizing enzymes, since these cells responded normally to the sterol metabolites 7alpha,27-dihydroxy-4-cholesten-3-one, 7alpha, 25-dihydroxy-4-cholesten-3-one, and 27-hydroxy-7-oxo-cholesterol. These metabolites, which all possessed an oxo group with a conjugated double bond in the steroid nucleus and a hydroxyl group in the side chain, did not seem to require further metabolism in order to be active. An impaired response to LDL was also seen in other human tumor cells, including breast carcinoma, colonic carcinoma, and malignant melanoma cells. Common to all the malignant cells was an intracellular shortage of 7alpha, 27-dihydroxy-4-cholesten-3-one caused by a decreased formation or an increased metabolism.

Cell Line, Transformed↗

PubMed1995

Low density lipoprotein (LDL) cholesterol is converted to 27-hydroxycholesterol in human fibroblasts. Evidence that 27-hydroxycholesterol can be an important intracellular mediator between LDL and the suppression of cholesterol production.

The formation of oxysterols in cultured human fibroblasts and their physiological roles as intracellular regulators of cholesterol production have been investigated. In the presence of low density lipoproteins (LDL) normal fibroblasts converted LDL cholesterol to the oxysterol 27-hydroxycholesterol in quantities apparently sufficient to down-regulate 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. The production of 27-hydroxycholesterol started 3-8 h after the addition of LDL to the incubation medium, and during this time the activity of HMG-CoA reductase decreased by 73%. Formation of other biologically active oxysterols such as 7 alpha-hydroxycholesterol, 24-hydroxycholesterol, and 25-hydroxycholesterol was not observed. When 27-hydroxylation of LDL cholesterol in mitochondria was selectively prevented by treating normal fibroblasts with cyclosporin A or by using fibroblasts genetically lacking sterol 27-hydroxylase, the suppressive effect of LDL on HMG-CoA reductase was reduced by a factor of about 10. In the absence of LDL or when the fibroblasts lacked LDL-receptors, the cells did not produce detectable amounts of 27-hydroxycholesterol, and HMG-CoA reductase was up-regulated. The results provide strong evidence that 27-hydroxycholesterol is an important intracellular mediator between LDL and the suppression of HMG-CoA reductase in human fibroblasts. The mitochondrial uptake/metabolism of LDL cholesterol seems to be a biologically important extension of the LDL pathway in human cells, since the mitochondrial products of LDL cholesterol may regulate cholesterol homeostasis or be precursors of steroid hormones or bile acids. This pathway, which has now been demonstrated in fibroblasts, may link together the two 22-year-old observations that LDL as well as oxysterols can down-regulate HMG-CoA reductase in cells.

Carbon Radioisotopes↗

PubMed1995

Structural specificity in the suppression of HMG-CoA reductase in human fibroblasts by intermediates in bile acid biosynthesis.

The effect of bile acid precursors on the activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was investigated. Cholesterol and 34 of its derivatives, including 23 potential intermediates in bile acid biosynthesis, were incubated with cultures of human fibroblasts for 24 h in the absence or presence of lipoproteins, and the activity of HMG-CoA reductase was then determined. In the absence of lipoproteins, many of the bile acid intermediates were inhibitory at a high concentration (2.5 microM), while only three, 27-hydroxycholesterol, 7 alpha, 27-dihydroxy-4-cholesten-3-one, and 7 alpha, 12 alpha, 27-trihydroxy-4-cholesten-3-one, caused a significant suppression at lower concentrations (often > 80% suppression at 0.25 microM). Even at 0.06 microM these sterols caused > 50% suppression of the enzyme activity. In addition, 27-hydroxy-4-cholesten-3-one, not usually considered to be an intermediate in bile acid biosynthesis, was a very potent inhibitor. Comparative studies showed that the effect of the three bile acid precursors was similar to that of 25-hydroxy-, 24-hydroxy-, and 7-oxo-cholesterol and 3 beta-hydroxy-5 alpha-cholest-8(14)-en-15-one. The presence of lipoproteins decreased or eliminated the inhibitory effect of most intermediates. Studies of the metabolism of the three most potent inhibitors in the fibroblasts indicated that the suppression was due to the compounds per se and not to products of their metabolism. The results show that a few specific intermediates in the formation of bile acids are potent suppressors of HMG-CoA reductase.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile Acids and Salts↗

PubMed1993

Synthesis of potential C27-intermediates in bile acid biosynthesis and their deuterium-labeled analogs.

In connection with studies of alternative pathways in bile acid biosynthesis, potential intermediates in a pathway starting with 27-hydroxylation of cholesterol have been prepared in natural and deuterated forms. Established methods were used to prepare 27-hydroxycholesterol and 3 beta-hydroxy-5-cholestenoic acid. Clemmensen reduction of kryptogenin in unlabeled and deuterated solvents yielded 27-hydroxy-cholesterol and 16-oxo-5-cholestene-3 beta,27-diol, which were separated by adsorption chromatography on Unisil. The labeled 27-hydroxycholesterol and 3 beta-hydroxy-5-cholestenoic acid derived from it consisted of molecules with seven (50%), six (20%), and eight (20%) deuterium atoms, and unlabeled molecules were not detected. The acetates of 27-hydroxycholesterol and methyl 3 beta-hydroxy-5-cholestenoate were 7 alpha-hydroxylated in a copper-catalyzed reaction with tert-butylperbenzoate, and the products were purified by chromatography on Unisil. The 7 beta-epimers were obtained as side products. Labeled 3 beta,7 alpha-dihydroxy-5-cholenoic acid was prepared in the same way from 3 beta-hydroxy-5-[2,2,4,4,23-2H5]-cholenoic acid. The 3-oxo-delta 4 analogs of the 3 beta-hydroxy-delta 5 compounds were prepared by oxidation with cholesterol oxidase. The labeled products had the same isotopic composition as the starting materials. Gas chromatographic retention indices and mass spectral characteristics of the trimethylsilyl ether derivatives of the neutral steroids and the methylated acids are given for all compounds.

Bile Acids and Salts↗

PubMed1993

Serum 7 alpha-hydroxy-4-cholesten-3-one concentrations in the evaluation of bile acid malabsorption in patients with diarrhoea: correlation to SeHCAT test.

The synthesis of bile acids is regulated by a homeostatic mechanism in which bile acids returning to the liver from the intestine inhibit their own synthesis. Serum concentrations of the bile acid intermediate 7 alpha-hydroxy-4-cholesten-3-one reflect the rate of bile acid synthesis whereas bile acid malabsorption can be determined by the SeHCAT test. This study was done to evaluate the correlation between the two tests in humans. Twenty eight patients with chronic diarrhoea were included in the study. Fasting serum was collected for the determination of 7 alpha-hydroxy-4-cholesten-3-one, and on the same day the gamma emitting bile acid analogue SeHCAT was given orally and its fractional catabolic rate assessed by repeated external counting over the upper abdomen during the next seven days. There was a highly significant positive correlation between the two tests (Rs = 0.80, p < 0.001). The results show a close relation between intestinal loss and hepatic synthesis of bile acids and imply that analysis of 7 alpha-hydroxy-4-cholesten-3-one in serum should now be evaluated as a possible convenient method for assessing bile acid malabsorption in patients with diarrhoea.

Adult↗

PubMed1993

Formation of 7 alpha- and 7 beta-hydroxylated bile acid precursors from 27-hydroxycholesterol in human liver microsomes and mitochondria.

In a search for enzymes involved in the formation of bile acids from 27-hydroxycholesterol in humans, the metabolism of this and other side-chain oxygenated steroids was studied in human liver microsomes and mitochondria. The microsomal fraction contained enzyme(s) catalyzing 7 alpha-hydroxylation of 27-hydroxycholesterol and 3 beta-hydroxy-5-cholestenoic acid, whereas the 7 alpha-hydroxylation of cholesterol and 3 beta-hydroxy-5-cholenoic acid was low. Only small amounts of 7 beta-hydroxylated products were formed. Purification and subfractionation of microsomal protein yielded a fraction of cytochrome P-450, which required NADPH and NADPH-cytochrome P-450 reductase and catalyzed 7 alpha-hydroxylation of the side-chain oxygenated 3 beta-hydroxy-delta 5-C27-steroids but was inactive toward cholesterol. Added cholesterol did not inhibit the observed enzymatic activity. The results provide evidence that this enzyme is different from cholesterol 7 alpha-hydroxylase. The mitochondrial fraction contained enzyme(s) that catalyzed an isocitrate-dependent 7 alpha-hydroxylation of 3 beta-hydroxy-5-cholestenoic acid. The activity was much lower with 27-hydroxycholesterol. The mitochondrial fraction also catalyzed the oxidation of the 27-hydroxy group and contained a 3 beta-hydroxy-delta 5-steroid dehydrogenase active on 7 alpha-hydroxylated C27-steroids. The metabolic end product of the reactions catalyzed by these enzymes was 7 alpha-hydroxy-3-oxo-4-cholestenoic acid. A considerable fraction of the 7 alpha-hydroxy-delta 5 intermediates was also converted to the corresponding 7 beta-hydroxysteroids, probably by way of the 7-oxosteroids, suggesting the presence of an epimerizing enzyme in the mitochondrial fraction.

Adult↗

PubMed1992

Cholesterol is converted to 7 alpha-hydroxy-3-oxo-4-cholestenoic acid in liver mitochondria. Evidence for a mitochondrial sterol 7 alpha-hydroxylase.

The metabolism of cholesterol in isolated intact pig liver mitochondria has been investigated. Six major cholesterol metabolites were identified by gas-liquid chromatography-mass spectrometry, the metabolic end product being 7 alpha-hydroxy-3-oxo-4-cholestenoic acid. Incubations with the synthesized intermediates suggested that the major pathway from cholesterol to this acid proceeds via the sequence of 26-hydroxylation, 7 alpha-hydroxylation, further oxidation of the side chain and oxidation/isomerization in the A-ring. The observed reactions prove that in addition to a sterol 26-hydroxylase, pig liver mitochondria contain significant amounts of a 7 alpha-hydroxylase active on side chain oxygenated 3 beta-hydroxy-delta 5-C27 steroids, an oxidoreductase active in the side chain of 26-hydroxylated steroids and a 3 beta-hydroxy-delta 5 steroid oxidoreductase active on 7 alpha-hydroxylated C27 steroids. Since 7 alpha-hydroxy-3-oxo-4-cholestenoic acid is believed to be an important precursor of chenodeoxycholic acid, this study shows that the first reactions in the biosynthesis of bile acids can be exclusively mitochondrial and thereby bypass microsomal cholesterol 7 alpha-hydroxylase as the rate-limiting enzyme.

Animals↗

PubMed1992

7 alpha-hydroxylation of 26-hydroxycholesterol, 3 beta-hydroxy-5-cholestenoic acid and 3 beta-hydroxy-5-cholenoic acid by cytochrome P-450 in pig liver microsomes.

Pig liver microsomes were found to catalyze the 7 alpha-hydroxylation of several potential bile acid precursors besides cholesterol. 26-Hydroxycholesterol, 3 beta-hydroxy-5-cholestenoic acid and 3 beta-hydroxy-5-cholenoic acid were all efficiently converted into the 7 alpha-hydroxylated products. Two cytochrome P-450 fractions showing 7 alpha-hydroxylase activity could be isolated. One fraction catalyzed 7 alpha-hydroxylation of 26-hydroxycholesterol, 3 beta-hydroxy-5-cholestenoic acid and 3 beta-hydroxy-5-cholenoic acid but was inactive towards cholesterol. The other fraction catalyzed 7 alpha-hydroxylation of cholesterol in addition to the other substrates. 26-Hydroxycholesterol in equimolar concentration did not inhibit the cholesterol 7 alpha-hydroxylase activity of this fraction. It is concluded that liver microsomes contain a cytochrome P-450 catalyzing 7 alpha-hydroxylation of 26-hydroxycholesterol, 3 beta-hydroxy-5-cholestenoic acid and 3 beta-hydroxy-5-cholenoic acid. The results indicate that this cytochrome P-450 is different from that catalyzing 7 alpha-hydroxylation of cholesterol.

Animals↗

PubMed1991

Bile acid synthesis in cultured human hepatoblastoma cells.

Biosynthetic pathways to bile acids have been studied in HepG2 cells, a well-differentiated human hepatoblastoma cell line. Cholesterol metabolites, in total 29, were isolated from culture media and cells by liquid-solid extraction and anion-exchange chromatography and were identified by gas-liquid chromatography-mass spectrometry. The production rates/concentrations of cholic acid (CA) and chenodeoxycholic acid (CDCA) in media from control cells were 71 and 74 ng/10(7) cells/h, respectively. Major bile acid precursors were 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA), 7 alpha, 12 alpha-dihydroxy-3-oxo-4-cholestenoic acid, 7 alpha-hydroxy-3-oxo-4-cholenoic acid, and 7 alpha, 12 alpha-dihydroxy-3-oxo-5 beta-cholanoic acid, their concentrations being 60, 30, 23, and 10 ng/10(7) cells/h, respectively. These and nine other isolated intermediates formed essentially complete metabolic sequences from cholesterol to CA and CDCA. The remaining steroids were metabolites of the intermediates or autooxidation products of cholesterol. These findings and the observed effect of dexamethasone on production rates suggest that in HepG2 cells the major biosynthetic pathways to primary bile acids start with 7 alpha-hydroxylation of cholesterol and oxidation to 7 alpha-hydroxy-4-cholesten-3-one followed by hydroxylation at either the 26 or 12 alpha position. CDCA is formed by the sequence of 26-hydroxylation, oxidation, and degradation of the side chain and A-ring reduction. CA is formed by the sequence of 12 alpha-hydroxylation, 26-hydroxylation, oxidation, and degradation of the side chain and reduction of the A-ring. An alternative pathway to CA included A-ring reduction of the intermediate 7 alpha, 12 alpha-dihydroxy-3-oxo-4-cholestenoic acid to form THCA prior to side chain cleavage. These pathways are not limited to HepG2 cells but may also be important in humans.

Bile Acids and Salts↗

PubMed1991

The plasma level of 7 alpha-hydroxy-4-cholesten-3-one reflects the activity of hepatic cholesterol 7 alpha-hydroxylase in man.

Circulating levels of 7 alpha-hydroxy-4-cholesten-3-one have been compared with activities of the rate-limiting enzyme in bile acid synthesis, microsomal cholesterol 7 alpha-hydroxylase, measured in liver biopsies obtained from patients undergoing surgery for gallstone disease. Some patients were treated with cholestyramine or bile acids prior to operation in order to alter the feed-back inhibition of the enzyme. The levels of the sterol were similar in untreated patients and in patients treated with ursodeoxycholic acid (median concentration 17 and 13 ng/ml, respectively), and so were the activities of the enzyme (median activity 7.0 and 5.5 pmol/min/mg protein, respectively). The sterol levels and enzyme activities were significantly increased in patients treated with cholestyramine (91 ng/ml and 45 pmol/min/mg protein) and decreased in patients treated with chenodeoxycholic acid (less than 2.0 ng/ml and 0.7 pmol/min/mg protein). There was a strong positive correlation (r = 0.90, P less than 0.00001) between levels of 7 alpha-hydroxy-4-cholesten-3-one in plasma and the activities of cholesterol 7 alpha-hydroxylase in the whole patient group. The results show that analysis of 7 alpha-hydroxy-4-cholesten-3-one in plasma is a sensitive and convenient method to determine relative rates of bile acid production in man.

Adult↗

PubMed1991

Ethanol has an acute effect on bile acid biosynthesis in man.

A single dose of ethanol, 0.4 g/kg body weight, was found to give a 5-15 fold increase of the plasma concentrations of 7 alpha-hydroxy-cholesterol and 7 alpha-hydroxy-4-cholesten-3-one in humans. The rise was maximal 4 h after ethanol ingestion, was dose-dependent and was not seen in a cholecystectomized subject. The effect was selective for these and some other 7 alpha-hydroxylated C27-intermediates in bile acid biosynthesis. The changes are compatible with an acute stimulation of cholesterol 7 alpha-hydroxylase possibly due to an ethanol-induced inhibition of gallbladder contraction resulting in an interruption of the enterohepatic circulation of bile acids. The effect is of interest in relation to the influence of ethanol consumption on cardiovascular and gallstone diseases.

Adult↗

PubMed1991

Occurrence of isomeric dehydrocholesterols in human plasma.

Three isomeric dehydrocholesterols were found in plasma from healthy subjects and patients with abnormal production or metabolism of cholesterol. These chemically labile steroids were isolated by a mild liquid-solid extraction procedure using octadecylsilane-bonded silica as sorbent. Sterol-protein interactions were minimized by diluting plasma with aqueous isopropanol. The dehydrocholesterols were identified by high-performance liquid chromatography-ultraviolet spectroscopy and gas chromatography-mass spectrometry as cholesta-5,7-dien-3 beta-ol (7-dehydrocholesterol), 5 alpha-cholesta-6,8(9)-dien-3 beta-ol (isodehydrocholesterol), and tentatively as cholesta-5,8(9)-dien-3 beta-ol. There was a strong positive correlation between plasma levels of the two former compounds, isodehydrocholesterol levels usually being about 1.4 times higher than those of 7-dehydrocholesterol. The median concentration of 7-dehydrocholesterol in plasma from healthy subjects was 52 ng/ml. Similar concentrations were found in colectomized patients (median concentration 47 ng/ml) and patients with extrahepatic cholestasis and alcoholic liver cirrhosis (median concentrations 79 and 67 ng/ml, respectively). Patients with ileal resection or under treatment with cholestyramine had elevated levels (median concentrations 142 and 160 ng/ml, respectively) whereas patients with primary biliary cirrhosis had subnormal levels (median concentration 26 ng/ml). The results are consistent with a positive correlation between levels of the dehydrocholesterols in plasma and the rate of cholesterol synthesis. The sterols were also analyzed in human skin and bile and the results indicate that the liver may be an important source of isodehydrocholesterol.

Adult↗

PubMed1990

Potential bile acid precursors in plasma--possible indicators of biosynthetic pathways to cholic and chenodeoxycholic acids in man.

The plasma concentrations of 3 beta-hydroxy-5-cholestenoic acid, 3 beta,7 alpha-dihydroxy-5-cholestenoic acid and 7 alpha-hydroxy-3-oxo-4-cholestenoic acid have been compared with that of 7 alpha-hydroxy-4-cholesten-3-one in healthy subjects and in patients with an expected decrease or increase of the bile acid production. In controls and patients with liver disease, the level of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid was positively correlated to that of 3 beta,7 alpha-dihydroxy-5-cholestenoic acid and not to that of 7 alpha-hydroxy-4-cholesten-3-one. In patients with stimulated bile acid formation the levels of the acids were not correlated to each other but there was a significant positive correlation between the levels of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid and 7 alpha-hydroxy-4-cholesten-3-one. These findings indicate that the precursor of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid differs depending on the activity of cholesterol 7 alpha-hydroxylase. Since the activity of this enzyme is reflected by the level of 7 alpha-hydroxy-4-cholesten-3-one in plasma the findings are compatible with a formation of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid from 3 beta,7 alpha-dihydroxy-5-cholestenoic acid when the rate of bile acid formation is normal or reduced and from 7 alpha-hydroxy-4-cholesten-3-one under conditions of increased bile acid synthesis. In support of this interpretation, 7 alpha,26-dihydroxy-4-cholesten-3-one was identified at elevated levels in plasma from patients with ileal resection or treated with cholestyramine. The levels of 7 alpha,12 alpha-dihydroxy-4-cholesten-3-one were also higher than normal in these patients. Based on these findings and previous knowledge, a model is proposed for the biosynthesis of bile acids in man. Under normal conditions, two major pathways, one "neutral" and one "acidic" or "26-oxygenated", lead to the formation of cholic acid and chenodeoxycholic acid, respectively. These pathways are separately regulated. When the activity of cholesterol 7 alpha-hydroxylase is high, the "neutral" pathway is most important whereas the reverse is true when cholesterol 7 alpha-hydroxylase activity is low. In cases with enhanced activity of cholesterol 7 alpha-hydroxylase, the "neutral" pathway is connected to the "acidic" pathway via 7 alpha,26-dihydroxy-4-cholesten-3-one, whereas a flow from the acidic pathway to cholic acid appears to be of minor importance.

Adult↗

PubMed1990

Stable isotope studies on steroid metabolism and kinetics: sulfates of 3 alpha-hydroxy-5 alpha-pregnane derivatives in human pregnancy.

The metabolism and production rates of 3 alpha-hydroxy-5 alpha-pregnan-20-one sulfate and the 3-sulfate and 3,20-disulfate of 5 alpha-pregnane-3 alpha,20 alpha-diol in pregnant women were studied. The steroid sulfates were labeled with deuterium in the 3 beta,11,11- or 3 beta,11,11,20 beta-positions and were injected intravenously. The deuterium content of steroids in the monosulfate and disulfate fraction of plasma collected at different times after the injection was determined by capillary column gas chromatography/mass spectrometry. The injected steroid sulfates underwent oxidoreduction at C-20 and 16 alpha-hydroxylation. In addition, the 3-sulfate of 5 alpha-pregnane-3 alpha,20 alpha-diol became hydroxylated at C-21. The pregnanediol and pregnanetriol monosulfates were also converted to disulfates. No evidence was obtained for a metabolic sequence involving hydrolysis, oxidoreduction, and resulfation at the C-3 position. Production rates and rates of metabolic transformations were determined using different one- and two-pool models. The production rate of the pregnanolone/pregnanediol monosulfate couple was 0.08 to 0.5 mmol/24 h, the variability probably depending both on individual factors and stage of pregnancy. The half-life time for oxidation and reduction at C-20 was 0.1 to 0.4 hours, reduction being the faster process. The half-life time for the turnover of the steroid skeleton was 1.3 to 3.3 hours. The injected steroid monosulfates were 16 alpha-hydroxylated at a rate of 1 to 8 mumol/24 h. A significant fraction of these 16 alpha-hydroxylated steroid sulfates, 0.5 to 25 mumol/24 h, was formed from other, probably unconjugated, precursors. The 16 alpha-hydroxylated steroid monosulfates underwent rapid oxidoreduction at C-20. The 3-sulfate of 5 alpha-pregnane-3 alpha,20 alpha-diol was hydroxylated at C-21. The production rate of 5 alpha-pregnane-3 alpha,20 alpha,21-triol 3-sulfate was 8 to 36 mumol/24 h in four women and 180 mumol/24 h in one woman, and this steroid was not formed from other precursors to a significant extent. 5 alpha-Pregnane-3 alpha,20 alpha-diol disulfate was a metabolic end product accounting for a major part of the elimination of the steroids injected. Its half-life time was 1.4 to 2.8 hours. The results show that the formation of sulfated steroids with a 3 alpha-hydroxy-5 alpha configuration may account for 50% of the metabolism of progesterone in late pregnancy.

Aryl Hydrocarbon Hydroxylases↗

PubMed1989

Concentrations of cholestenoic acids in plasma from patients with liver disease.

The concentrations of 3 beta-hydroxy-5-cholestenoic acid, 3 beta,7 alpha-dihydroxy-5-cholestenoic acid, and 7 alpha-hydroxy-3-oxo-4-cholestenoic acid were determined in plasma from patients with different liver diseases and compared with those of unconjugated and conjugated C24 bile acids. The levels of the cholestenoic acids were similar in patients with extrahepatic cholestasis and in controls (median concentration 153 and 162 ng/ml, respectively), whereas significantly elevated levels were found in plasma from patients with primary biliary cirrhosis (median concentration 298 ng/ml) and alcoholic liver cirrhosis (median concentration 262 ng/ml). As expected, conjugated C24 bile acids were elevated in most patients whereas the corresponding unconjugated compounds were low in cholestasis and elevated in alcoholic liver cirrhosis. The levels of the individual C27 acids were usually positively correlated to each other and also to the levels of conjugated C24 bile acids in plasma from patients with liver cirrhosis. In contrast, there was no correlation between the levels of C27 acids and conjugated bile acids in patients with extrahepatic cholestasis. The levels of unconjugated C24 bile acids were not correlated to C27 acids or conjugated bile acids in any of the groups. The results indicate that there is a close metabolic relationship between the individual C27 acids, that they do not participate in an enterohepatic circulation, and that the liver is important for their elimination/metabolism.

Aged↗

PubMed1989

Concentrations of cholestenoic acids in plasma from patients with reduced intestinal reabsorption of bile acids.

The concentrations of 3 beta-hydroxy-5-cholestenoic acid, 3 beta,7 alpha-dihydroxy-5-cholestenoic acid, and 7 alpha-hydroxy-3-oxo-4-cholestenoic acid were determined in plasma from patients treated with cholestyramine or subjected to resection of the ileum or colon. The values were compared with those for conjugated and unconjugated C24 bile acids. Patients with an intact ileum but without colon had normal levels of cholestenoic acids. Patients treated with cholestyramine or with ileal resection had elevated levels of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid (median values 189 and 233 ng/ml, respectively, compared to 85 ng/ml in controls). The levels of the other two C27 acids were normal in cholestyramine-treated and low in ileoresected patients and were positively correlated to each other but not to the 3-oxo-delta 4 acid. There were no consistent correlations between the levels of C27 acids and those of conjugated or unconjugated C24 bile acids. The results indicate an increased formation of 7 alpha-hydroxy-3-oxo-4-cholestenoic acid in subjects having a stimulated activity of cholesterol 7 alpha-hydroxylase.

Adult↗

PubMed1988

Levels of 7 alpha-hydroxy-4-cholesten-3-one in plasma reflect rates of bile acid synthesis in man.

A method for analysis of 7 alpha-hydroxy-4-cholesten-3-one in plasma is described. Following solid-phase extraction/purification the compound is determined by high-performance liquid chromatography using a UV detector. The median concentration in healthy subjects was 12 ng/ml (range 3-40). The levels were lower in diseases associated with a low bile acid production: extrahepatic cholestasis, less than 1.5 ng/ml (range less than 0.9-3); liver cirrhosis less than 1.5 ng/ml (range less than 0.9-38), and higher in diseases associated with a high bile acid production: cholestyramine treatment, 188 ng/ml (range 54-477); ileal resection 397 ng/ml (range 128-750). The levels were essentially normal in patients with colon resection. The results are consistent with a strong positive correlation between the levels of 7 alpha-hydroxy-4-cholesten-3-one in plasma and the rate of bile acid synthesis.

Adult↗

PubMed1988

Vitamin D metabolism in human pregnancy. Concentrations of free and sulphated 25-hydroxyvitamin D3 in maternal and fetal plasma at term.

The concentrations of free and sulphated 25-hydroxyvitamin D3 in 20 paired maternal-cord plasma samples obtained at delivery have been determined. The compounds were isolated by liquid-solid extraction at elevated temperature, and the sulphate was purified by anion exchange chromatography prior to hydrolysis and analysis by high-performance liquid chromatography. The study shows that unconjugated 25-hydroxyvitamin D3 is predominant in maternal plasma (mean 20 ng/ml) whereas the sulphate is the major form of vitamin D3 in fetal circulation (mean 21 ng/ml plasma). The total concentration of the two compounds in cord plasma (mean 35 ng/ml) was significantly higher than that in maternal plasma (mean 30 ng/ml). Positive correlations were obtained between maternal and cord plasma levels of free 25-hydroxyvitamin D3, between maternal and cord plasma levels of sulphated 25-hydroxyvitamin D3 and between plasma levels of the maternal free compound and the fetal sulphate. There was also a relationship between the levels of free and sulphated 25-hydroxyvitamin D3 in cord plasma. The results suggest that sulphation may be a physiologically important reaction for deactivating and/or trapping secosteroids in the fetus.

Adult↗