RESUMO
Bile acids and bile alcohols in the form of their conjugates are amphipathic end products of cholesterol metabolism with multiple physiological functions. The great variety of bile acids and bile alcohols that are present in vertebrates are tabulated. Bile salts have an enterohepatic circulation resulting from efficient vectorial transport of bile salts through the hepatocyte and the ileal enterocyte; such transport leads to the accumulation of a pool of bile salts that cycles between the liver and intestine. Bile salt anions promote lipid absorption, enhance tryptic cleavage of dietary proteins, and have antimicrobial effects. Bile salts are signaling molecules, activating nuclear receptors in the hepatocyte and ileal enterocyte, as well as an increasing number of G-protein coupled receptors. Bile acids are used therapeutically to correct deficiency states, to decrease the cholesterol saturation of bile, or to decrease the cytotoxicity of retained bile acids in cholestatic liver disease.
Assuntos
Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/metabolismo , Doenças Biliares/patologia , Doenças Biliares/terapia , Animais , Doenças Biliares/fisiopatologia , Transporte Biológico , Circulação Sanguínea , Proteínas de Transporte/metabolismo , Circulação Êntero-Hepática , Humanos , Glicoproteínas de Membrana/metabolismoRESUMO
5alpha-Cyprinol sulfate was isolated from bile of the Asiatic carp, Cyprinus carpio. 5alpha-Cyprinol sulfate was surface active and formed micelles; its critical micellization concentration (CMC) in 0.15 M Na+ using the maximum bubble pressure device was 1.5 mM; by dye solubilization, its CMC was approximately 4 mM. At concentrations >1 mM, 5alpha-cyprinol sulfate solubilized monooleylglycerol efficiently (2.1 molecules per mol micellar bile salt). When infused intravenously into the anesthetized rat, 5alpha-cyprinol sulfate was hemolytic, cholestatic, and toxic. In the isolated rat liver, it underwent little biotransformation and was poorly transported (Tmax congruent with 0.5 micromol/min/kg) as compared with taurocholate. 5alpha-Cyprinol, its bile alcohol moiety, was oxidized to its corresponding C27 bile acid and to allocholic acid (the latter was then conjugated with taurine); these metabolites were efficiently transported. 5alpha-Cyprinol sulfate inhibited taurocholate uptake in COS-7 cells transfected with rat asbt, the apical bile salt transporter of the ileal enterocyte. 5alpha-Cyprinol had limited aqueous solubility (0.3 mM) and was poorly absorbed from the perfused rat jejunum or ileum. Sampling of carp intestinal content indicated that 5alpha-cyprinol sulfate was present at micellar concentrations, and that it did not undergo hydrolysis during intestinal transit. These studies indicate that 5alpha-cyprinol sulfate is an excellent digestive detergent and suggest that a micellar phase is present during digestion in cyprinid fish.
Assuntos
Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/metabolismo , Colestanóis/química , Colestanóis/metabolismo , Animais , Bile/química , Ácidos e Sais Biliares/isolamento & purificação , Ácidos e Sais Biliares/toxicidade , Transporte Biológico , Biotransformação , Carpas/metabolismo , Linhagem Celular , Colestanóis/isolamento & purificação , Colestanóis/toxicidade , Técnicas In Vitro , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Estrutura Molecular , Perfusão , Ratos , Espectrometria de Massas por Ionização por Electrospray , Tensão SuperficialRESUMO
Urinary androstanes from seven species of male great apes (human, bonobo, chimpanzee, lowland gorilla, mountain gorilla, Bornean orangutan, and Sumatran orangutan) were separated by HPLC and detected by RIA using two testosterone antibodies. All animals examined showed the presence of testosterone and six additional immunoreactive peaks. Although testosterone was the dominant peak (85%) in human urine, its proportion in urine was much less in the other apes, ranging from a high of 59% in the bonobo and chimpanzee to a low of 24% in the mountain gorilla. Urinary androstanes were also directly visualized using nano-spray mass spectrometry (nanoESI-MS). Although the RIA can qualitatively produce a strong signal for testosterone in unchromatographed urine, it is quantitatively present only as a trace metabolite, as demonstrated by nanoESI-MS. The combination of the two techniques showed large differences in androstane metabolism between the seven species. A previously undescribed testosterone metabolite (tentatively identified as either delta1- or delta6-testosterone sulfate) was present in significant proportions in all of the non-human apes examined. We conclude that in the great apes, testosterone is only a trace metabolite in urine, and as a consequence, its measurement may not produce results that parallel the levels of serum testosterone. The RIA measurement of urinary testosterone in part records additional androstane metabolites, which vary even between closely related genera, making the results neither equivalent with nor comparable to different species.
Assuntos
Androstanos/urina , Hominidae/urina , Animais , Cromatografia Líquida de Alta Pressão , Desidroepiandrosterona/urina , Gorilla gorilla/urina , Humanos , Masculino , Pan paniscus/urina , Pan troglodytes/urina , Pongo pygmaeus/urina , Radioimunoensaio , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray , Testosterona/sangue , Testosterona/urinaRESUMO
The Shoebill stork, an enigma phylogenetically, was found to contain as its dominant biliary bile acid 16alpha-hydroxychenodeoxycholic acid, a heretofore undescribed bile acid. The bile acid occurred as its taurine N-acyl amidate; structure was established by nuclear magnetic resonance (NMR) and mass spectrometry (MS). A search for this novel bile acid in other Ciconiiformes showed that it constituted >92% of biliary bile acids in five of nine herons in the Ardidae, but was absent in all other families (Ciconiidae, Threskiornithidae, Scopidae, Phoenicopteridae). The presence of this biochemical trait in the Shoebill stork and certain herons suggests that these birds are closely related.
Assuntos
Ácidos e Sais Biliares/análise , Aves/fisiologia , Ácido Quenodesoxicólico/análogos & derivados , Ácido Quenodesoxicólico/análise , Ácido Quenodesoxicólico/química , Filogenia , Animais , Ácidos e Sais Biliares/química , Aves/classificação , Ácido Quenodesoxicólico/isolamento & purificação , Cromatografia Gasosa-Espectrometria de Massas/métodos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Especificidade da EspécieRESUMO
Fecal extracts from a pregnant black rhinoceros, Diceros bicornis, were analyzed by radioimmunoassay, HPLC, and by GC-mass spectrometry. From 40 g of dried feces a total of 33 pregnanes in the C(21)O(2) series, including a number of novel 17 alpha epimers were identified. No progesterone was recovered. The principal progesterone metabolite by mass was 5 alpha-pregnan-3 beta,20 alpha-diol (44.5%), which did not cross react with the antibody used in our RIA. The antibody recognized progesterone and pregnanes with 20-one configuration, which when combined made up less than 15% of the total C(21)O(2) steroid mass. Of the 33 pregnanes in the C(21)O(2) series identified, 81%, by mass, were in the 5 alpha-configuration. These results are compared with studies in other rhinoceros species (Asian and Sumatran) in which pregnanes in the 5 beta-configuration are the major fecal metabolites, and the white rhinoceros in which pregnanes in the 5 alpha-configuration are the dominant form.
Assuntos
Fezes/química , Perissodáctilos/metabolismo , Pregnanos/química , Pregnanos/isolamento & purificação , Animais , Especificidade de Anticorpos , Cromatografia Líquida de Alta Pressão , Reações Cruzadas , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Gravidez , Radioimunoensaio , Fatores de TempoRESUMO
Bile acid malabsorption is often present in patients after near-total proctocolectomy and ileal pouch-anal canal anastomosis, suggesting ileal dysfunction. Experiments were performed in dogs to compare bile acid absorption after a modified procedure, in which a jejunal pouch was interposed between the terminal ileum and the distal rectum, with that after a conventional ileal pouch operation. Fecal bile acid output (equivalent to hepatic bile acid biosynthesis) and composition were determined by gas chromatography/mass spectrometry in five jejunal pouch dogs and in five ileal pouch dogs more than 6 months after operation. Fecal bile acid output in the jejunal pouch dogs (mean +/- standard deviation) was 215 +/- 59 mg/day (10.1 +/- 2.7 mg/kg-day), a value similar to that obtained in the ileal pouch dogs (261 +/- 46 mg/day [12.8 +/- 3.1 mg/kg-day]; P >0.05). These values were also similar to those reported by others for healthy unoperated dogs, indicating that increased bile acid biosynthesis occurring in response to bile acid malabsorption was not present. Fecal bile acids in pouch dogs were completely deconjugated and extensively 7-dehydroxylated (jejunal pouch = 90.4% dehydroxylated; ileal pouch = 88.6% +/- 6.6% dehydroxylated) and consisted predominantly of deoxycholic acid derivatives. We conclude that when either a jejunal pouch or an ileal pouch is used as a rectal substitute in dogs, an anaerobic pouch flora develops that efficiently deconjugates and dehydroxylates bile acids, rendering them membrane permeable. The resultant passive absorption of unconjugated bile acids appears to compensate for any loss of active ileal absorption of conjugated bile acids, and bile acid malabsorption does not occur.
Assuntos
Ácidos e Sais Biliares/metabolismo , Proctocolectomia Restauradora , Absorção , Animais , Cães , Fezes/química , Cromatografia Gasosa-Espectrometria de Massas , Reto/cirurgiaRESUMO
Urinary and fecal hormones were analyzed on average every other day in 17 female bonobos kept at four US zoos (San Diego Zoo and Wild Animal Park, Milwaukee, Columbus, and Cincinnati). Ovarian cycle activity was monitored throughout the 15-month study period using estrogen and progesterone profiles and swelling charts. Behavioral data on sexual activity were also collected on a daily basis. Fecal and urinary samples were analyzed using high pressure liquid chromatography (HPLC), gas chromatography-mass spectrometry (CG-MS), and nanoelectrospray. Preliminary results indicate that in urine, both conjugated progestin and estrogen metabolites were abundant, while in fecal samples, free progestin metabolites from the 5a-pregnane series were found. Although traces of estrogen metabolites were detected in fecal samples, long-term monitoring of ovarian activity in our study yielded no meaningful estrogen profiles. In contrast, fecal progestin profiles, after adjusting for a one-day delay in excretion, closely matched the corresponding urinary progestin profiles. Using the identical antibody and tracer for both, fecal and urinary progestins, fecal samples yielded approximately ten times the relative amount of progestins compared to urinary progestins. Thus, when converted using a regression formula, fecal progestins may complete the picture obtained from urinary progestins, particularly in cases where the urine sample record is unavailable or incomplete. Evidence of the usefulness of urinary cortisol as a measure of stress is presented.
RESUMO
BACKGROUND & AIMS: Dihydroxy bile acids induce a bicarbonate-rich hypercholeresis when secreted into canalicular bile in unconjugated form; the mechanism is cholehepatic shunting. The aim of this study was to identify a xenobiotic that induces hypercholeresis by a similar mechanism. METHODS: Five organic acids (sulindac, ibuprofen, ketoprofen, diclofenac, and norfloxacin) were infused into rats with biliary fistulas. Biliary recovery, bile flow, and biliary bicarbonate were analyzed. Sulindac transport was further characterized using Tr(-) rats (deficient in mrp2, a canalicular transporter for organic anions), the isolated perfused rat liver, and hepatocyte membrane fractions. RESULTS: In biliary fistula rats, sulindac was recovered in bile in unconjugated form and induced hypercholeresis of canalicular origin. Other compounds underwent glucuronidation and were not hypercholeretic. In the isolated liver, sulindac had delayed biliary recovery and induced prolonged choleresis, consistent with a cholehepatic circulation. Sulindac was secreted normally in Tr(-) rats, indicating that its canalicular transport did not require mrp2. In the perfused liver, sulindac inhibited cholyltaurine uptake, and when coinfused with cholyltaurine, induced acute cholestasis. With both basolateral and canalicular membrane fractions, sulindac inhibited cholyltaurine transport competitively. CONCLUSIONS: Sulindac is secreted into bile in unconjugated form by a canalicular bile acid transporter and is absorbed by cholangiocytes, inducing hypercholeresis. At high flux rates, sulindac competitively inhibits canalicular bile salt transport; such inhibition may contribute to the propensity of sulindac to induce cholestasis in patients.
Assuntos
Anti-Inflamatórios não Esteroides/metabolismo , Canalículos Biliares/metabolismo , Ductos Biliares/irrigação sanguínea , Bile/metabolismo , Circulação Hepática , Sulindaco/metabolismo , Animais , Anti-Inflamatórios não Esteroides/sangue , Bicarbonatos/metabolismo , Bile/fisiologia , Fístula Biliar/metabolismo , Transporte Biológico/efeitos dos fármacos , Biotransformação , Técnicas In Vitro , Cinética , Masculino , Perfusão , Ratos , Ratos Sprague-Dawley , Sulindaco/antagonistas & inibidores , Sulindaco/sangue , Sulindaco/farmacocinética , Ácido Taurocólico/metabolismoRESUMO
BACKGROUND & AIMS: Although fat malabsorption in the short-bowel syndrome is caused in part by decreased bile acid secretion, bile acid replacement therapy is not used because of the belief that ingested bile acids would worsen diarrhea, outweighing the benefits of improved fat absorption. This study compared the effect of a natural conjugated bile acid mixture from ox bile with that of cholylsarcosine, a synthetic conjugated bile acid, on fat absorption and diarrhea in a patient with the short-bowel syndrome. Cholylsarcosine is resistant to bacterial metabolism and has no cathartic activity. METHODS: Metabolic balance studies and a clinical trial were performed in an emaciated patient with the short-bowel syndrome and ileostomy in whom parenteral nutrition could not be used. RESULTS: In balance studies, conjugated bile acid replacement therapy with either preparation caused fat absorption to increase by approximately 40 g/day. Calcium absorption also increased. Neither bile acid product caused a clinically significant increase in ileostomy water output. During a 4-month outpatient trial, while the patient ingested 2 g/meal natural bile acids, her weight increased from 80 to 98 lb, without side effects. CONCLUSIONS: Conjugated bile acid replacement therapy should be part of the armamentarium for the treatment of selected patients with the short-bowel syndrome.
Assuntos
Ácidos e Sais Biliares/uso terapêutico , Síndrome do Intestino Curto/tratamento farmacológico , Animais , Aspartato Aminotransferases/sangue , Ácidos e Sais Biliares/efeitos adversos , Peso Corporal/efeitos dos fármacos , Bovinos , Ácidos Cólicos/efeitos adversos , Ácidos Cólicos/uso terapêutico , Diarreia/tratamento farmacológico , Gorduras na Dieta/metabolismo , Ingestão de Energia , Fezes/química , Feminino , Seguimentos , Humanos , Ileostomia , Absorção Intestinal , Pessoa de Meia-Idade , Sarcosina/efeitos adversos , Sarcosina/análogos & derivados , Sarcosina/uso terapêutico , Síndrome do Intestino Curto/metabolismo , Síndrome do Intestino Curto/fisiopatologiaRESUMO
jj biliary bile acid composition of the adult and neonatal domestic rabbit, as well as that of the adult brush rabbit, was characterized. In adult domestic rabbits, the dominant bile acid present was deoxycholic acid (88% of total bile acids), a secondary bile acid formed by the bacterial 7-dehydroxylation of cholic acid. Although most of the bile acids present were conjugated with glycine, two exceptions were observed. About 3% of deoxycholic acid was conjugated, in N-acyl linkage, with glycyl-taurine. Chenodeoxycholic acid, which composed <1% of wile acids, was conjugated solely with taurine. The bile of neonatal rabbits contained a greater percentage of primary bile acids, and bile acids were conjugated to a much greater extent with taurine. The adult brush rabbit had a bile acid composition similar to that of the domestic rabbit, but about one-third of all bile acids were conjugated with taurine. In addition, lithocholic acid was present as its sulfated amidate, whereas in the domestic rabbit, lithocholic acid was conjugated solely with glycine. The biliary bile acid composition of rabbits appears to be unique both in terms of the predominant steroid moiety, as well as in the modes of conjugation.
Assuntos
Ácidos e Sais Biliares/química , Sistema Biliar/metabolismo , Ácido Desoxicólico/análogos & derivados , Coelhos/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Glicosilação , Masculino , Espectrometria de Massas , Taurina/análiseRESUMO
Estrone and estrone conjugates were measured in the feces of three female pygmy lorises (Nycticebus pygmaeus) throughout estrus, pregnancy, and the postpartum period. Two females gave birth to twins, while the third had a single stillborn. A comparison between the hormonal profiles of these three pregnancies with each other and with previously reported pregnancies resulting in singletons or twins [Jurke et al., American Journal of Primatology 41:103-115, 1997] revealed a characteristic pattern of hormonal excretion. This report adds data to and confirms previous claims that monitoring estrone in this species provides a tool to assess gestation length (via determination of estrus period) and to predict the date of parturition and the number of offspring. However, there was an exceptional case of a pregnancy which had the hormonal appearance of a twin pregnancy but resulted in a single stillborn infant. This case prompted us to search for new insights into the characteristics and the origin of the estrogens that are excreted into the feces. Aromatase activity was evaluated in five partial placentae.
Assuntos
Estrona/análise , Lorisidae/fisiologia , Testes de Gravidez/veterinária , Prenhez , Animais , Estrona/análogos & derivados , Fezes/química , Feminino , Idade Gestacional , Gravidez , GêmeosRESUMO
We postulated that coadministration of cholylsarcosine with ursodeoxycholic acid might provide additional benefit to primary biliary cirrhosis patients with an incomplete response to ursodeoxycholic acid. Our aim was to test the tolerability and the effect of adjuvant cholylsarcosine on liver tests and plasma cholesterol in primary biliary cirrhosis patients receiving ursodeoxycholic acid. Four primary biliary cirrhosis patients, who, despite more than a year of ursodeoxycholic acid therapy, had one or more liver tests persistently equal to or greater than twice the upper limit of normal, received cholylsarcosine (12-15 mg/kg/day) in addition to ursodeoxycholic acid (13-15 mg/kg/day) for six weeks in an open label study. Values of liver tests and plasma cholesterol, determined every two weeks, remained unchanged. One patient discontinued cholylsarcosine at week 4 because of new-onset pruritus. Analysis of duodenal bile acids in one patient showed 52% enrichment in cholylsarcosine and hydrophilic bile acids constituted 87% of total bile acids. It is concluded that the addition of cholylsarcosine to ursodeoxycholic acid did not influence liver tests in four primary biliary cirrhosis patients who had not responded completely to ursodeoxycholic acid alone. Cholylsarcosine was absorbed and became a dominant biliary bile acid; its administration was associated with increased pruritus.
Assuntos
Colagogos e Coleréticos/uso terapêutico , Ácidos Cólicos/uso terapêutico , Cirrose Hepática Biliar/tratamento farmacológico , Sarcosina/análogos & derivados , Ácido Ursodesoxicólico/uso terapêutico , Adulto , Colesterol/sangue , Quimioterapia Combinada , Feminino , Humanos , Cirrose Hepática Biliar/sangue , Cirrose Hepática Biliar/fisiopatologia , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , Sarcosina/uso terapêuticoRESUMO
Studies were performed to characterize hepatic and intestinal transport, as well as biotransformation during transport, of a spectrum of fluorescent bile acids containing a fluorophore attached to the side chain. The following two classes of compounds were studied: 1) aminofluorescein (amF) coupled directly to the carboxylic group of a bile acid which was cholic, ursodeoxycholic, or cholylglycine; and 2) nitrobenzoxadiazolyl (NBD) coupled to the epsilon-amino group of a lysine conjugated bile acid, which was cholic or ursodeoxycholic. Fluorescein, a cholephilic organic anion, was studied as a control. Fluorescent bile acids were synthesized and their structures confirmed by nuclear magnetic resonance and mass spectrometry. Using the biliary fistula rat, hepatic transport, biotransformation, and choleretic activity were defined; intestinal absorption was assessed by jejunal or ileal perfusion studies. All fluorescent bile acids had hepatic transport maxima about one-sixth that reported for cholyltaurine, but derivatives of cholylglycine were transported best. Bile acids underwent little (<5%) biotransformation during hepatocyte transport. Only the amF conjugate of cholylglycine had normal choleretic activity; other compounds were hypocholeretic or cholestatic. In contrast, fluorescein was well transported, was partly glucuronidated, and had normal choleretic activity. NBD-tagged, but not amF-tagged, bile acids were actively transported by the intestine (ileum > jejunum), and no fluorescent bile acid had passive intestinal permeability; NBD-tagged bile acids were biotransformed during intestinal transport (jejunum > ileum). We conclude that the structure of the fluorophore as well as that of the bile acid influences transport by the hepatocyte and enterocyte. These fluorescent bile acids differ from fluorescein in being impermeable to cell membranes and undergoing little biotransformation during hepatocyte transport. Of these fluorescent bile acids, cholylglycylamF has hepatocyte transport and choleretic properties most closely resembling those of a natural bile acid.
Assuntos
Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/farmacocinética , Fluorescência , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Animais , Bile/metabolismo , Ácidos e Sais Biliares/farmacologia , Transporte Biológico/fisiologia , Biotransformação/fisiologia , Colagogos e Coleréticos/farmacologia , Absorção Intestinal/fisiologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
To assess the effect of side chain length on the metabolism and physiological effects of homologues of chenodeoxycholic acid (CDCA), dinorCDCA, the C22 homologue, was synthesized and its hepatic biotransformation, transport kinetics, and choleretic properties were defined in rat and hamster biliary fistula and in isolated perfused rat liver. Results were compared with those of norCDCA, the C23 homologue, and of CDCA, the natural C24 homologue. In the rat, dinorCDCA was secreted mostly in unconjugated form (the majority as dinor-alpha-muricholic acid); the remainder was glucuronidated. In the hamster, glucuronidation was greater, and the unconjugated fraction contained equal parts of dinorCDCA and 5beta-hydroxy-dinorCDCA. NorCDCA was glucuronidated extensively (70%, rat; 40%, hamster). CDCA, in contrast, was efficiently amidated with taurine or glycine. In the perfused liver, the initial uptake rate of all three homologues was identical; later, regurgitation and/or cholehepatic shunting of dinorCDCA and norCDCA, but not of CDCA, occurred. In rats and hamsters with biliary fistulas, dinorCDCA and norCDCA, but not CDCA, induced a bicarbonate-rich hypercholeresis of canalicular origin. Hypercholeresis was not induced by the taurine conjugate of dinorCDCA. Hepatobiliary retention of both dinorCDCA and norCDCA occurred, consistent with efficient ductular absorption (calculated to be 94%) and cholehepatic cycling of the unmetabolized bile acids. It is concluded that dinorCDCA, as norCDCA, is inefficiently amidated, is metabolized as a xenobiotic, and induces hypercholeresis. DinorCDCA is the first dihydroxy bile acid to be identified that is secreted largely in unconjugated form in bile.
Assuntos
Ácido Quenodesoxicólico/farmacocinética , Colagogos e Coleréticos/farmacologia , Fígado/metabolismo , Animais , Fístula Biliar/metabolismo , Transporte Biológico , Biotransformação , Ácido Quenodesoxicólico/análogos & derivados , Ácido Quenodesoxicólico/farmacologia , Cricetinae , Ratos , Solubilidade , Relação Estrutura-AtividadeRESUMO
An improved method is described for the quantitation of lidocaine and its dominant metabolites in rat plasma, 3-hydroxy-lidocaine glucuronide and 3-hydroxy-MEG-X glucuronide. Frozen plasma samples (100-200 microliters) were thawed and deproteinated by precipitation with acetonitrile, before the conversion of glucuronidates into their respective hydroxylated forms by acid hydrolysis. After extraction with solid-phase C18 cartridge chromatography, the metabolites and parent drug were analyzed by capillary gas chromatography-nitrogen phosphorus detection, without derivativization. A detection limit of 0.005 microgram/ml for lidocaine and nonglucuronidated metabolites and 0.01 microgram/ml for glucuronidated metabolites was achieved. The method offers significant improvements in sensitivity relative to existing techniques, which should be of specific benefit to studies in which sample volume is limited, such as those concerned with the pharmacokinetics of lidocaine metabolism in small-animal pain state models.
Assuntos
Lidocaína/sangue , Lidocaína/metabolismo , Animais , Cromatografia em Camada Fina , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The hepatic biotransformation in the rat and hamster of 23(R)-hydroxychenodeoxycholic acid (23(R)OH-CDCA), the alpha-hydroxy derivative of CDCA, was defined; some physiological and physicochemical properties were also assessed. 23(R)OH-CDCA was isolated from duck bile; [24-14C]23(R)OH-CDCA was synthesized. The compound was administered intravenously to anesthetized biliary fistula rats at doses of 1, 3, or 5 mu mol/kg-min and to hamsters at 3 mu mol/min-kg. Biliary bile acids and radioactivity were analyzed by thin-layer chromatography (TLC), high pressure liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS). Recovery of radioactivity in bile was incomplete (50-70% of infused dose); some was also recovered as breath 14CO2. Radioactivity in bile was present as unchanged compound (25-50%, dose-dependent) and its conjugates (with taurine, with glycine, or with glucuronate). Nor-CDCA (C23) was present in bile (in both unconjugated and conjugated form), indicating that 23(R)OH-CDCA had undergone oxidative decarboxylation (alpha-oxidation) with loss of the C-24 carboxyl group. The alpha-oxidation was 20 +/- 5% (mean +/- SD) of administered dose in the rat and was not dose-dependent; in hamsters, alpha-oxidation was 35 +/- 8%. In rats, the S isomer of 23OH-CDCA also underwent alpha-oxidation (10 +/- 2%). Nor-CDCA also underwent 6beta-hydroxylation to form nor-alpha-muricholic acid, as well as reduction of its C-23 carboxyl group to form the C23 alcohol. The taurine conjugate of 23(R)OH-CDCA [23(R)OH-CDC-tau] was prepared synthetically and characterized by 1H-NMR. By surface tension measurements, it had a critical micellization concentration (CMC) of 3.5 mM (in 0.15 M Na+), as compared to 1.8 mM for CDC-taurine. Aqueous solubility of 23(R)OH-CDCA increased markedly above pH 5, compared to pH 7 for CDCA. When incubated with cholylglycine hydrolase, 23(R)OH-CDC-tau was deconjugated at a rate one-fourth that of CDC-tau. It is concluded that the presence of a 23(R)-hydroxyl group in a 3alpha, 7alpha-dihydroxy bile acid alters its metabolism in the rodent hepatocyte, as evidenced by inefficient conjugation with taurine or glycine, alpha-oxidation to nor (C23) bile acid, and reduction of the nor bile acid to the primary alcohol. The taurine conjugate of 23(R)OH-CDCA, a major biliary bile acid of marine mammals and wading birds, is a biological detergent with properties superior to those of the taurine conjugate of CDCA. Natural C23 nor-bile acids may be formed by alpha-oxidation of alpha-hydroxy C24 bile acids.
Assuntos
Ácido Quenodesoxicólico/análogos & derivados , Fígado/metabolismo , Animais , Bile/química , Bile/metabolismo , Transporte Biológico , Biotransformação , Testes Respiratórios , Ácido Quenodesoxicólico/química , Ácido Quenodesoxicólico/metabolismo , Cricetinae , Patos , Masculino , Mesocricetus , Oxirredução , Ratos , Ratos Sprague-Dawley , EstereoisomerismoRESUMO
The biliary bile acid composition of 30 species of pigeons and doves belonging to seven genera in the avian order Columbiformes was determined using TLC, HPLC, GLC/MS, LSIMS, and NMR. In 23 of 25 species of fruit pigeons and doves, chenodeoxycholic acid was the major bile acid (> 50%). In only 1 species (Ptilinopus ornatus) was cholic the major bile acid. A number of species (7 of 15 species in the genus Ptilinopus, and 6 of 9 species in the genus Ducula) contained 1 beta,3 alpha,7 alpha-trihydroxy-5 beta-cholan-24-oic acid in proportions ranging from 2 to 43%. This 1 beta-hydroxy derivative of chenodeoxycholic acid has not been previously identified as a major biliary bile acid in vertebrates. Five of 15 species of the genus Ptilinopus, 5 of 9 species of the genus Ducula, and the only species examined for the genus Gymnophaps contained 23R-hydroxy chenodeoxycholic acid in detectable proportions, ranging from 1 to 4%. Bile acids were conjugated (in N-acyl linkage) with glycine and taurine in 28 species and with only taurine in 2 species. The fruit pigeons are the first non-mammalian genera identified to date in whom bile acids are conjugated with glycine, as well as with taurine. An incidental finding was that a gallbladder was present in 3 genera (Ptilinopus, Ducula, and Gymnophaps) and absent in 4 genera (Gallicolumba, Chalcophaps, Otidiphaps, and Treron).
Assuntos
Ácidos e Sais Biliares/análise , Bile/química , Ácido Quenodesoxicólico/análogos & derivados , Columbidae , Glicina , Taurina , Animais , Ácido Quenodesoxicólico/análise , Ácido Quenodesoxicólico/química , Ácido Cólico , Ácidos Cólicos/análise , Cromatografia Líquida de Alta Pressão , Glicina/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Taurina/químicaRESUMO
Fresh and desiccated gall bladders of the Ursidae family (bears) obtained as criminal evidence were characterized by analysis of the principal biliary components, mainly ursodeoxycholyl-taurine, cholyl-taurine and chenodeoxycholyl-taurine using TLC and HPLC. This bile acids profile appears to be an Ursidae family characteristic. Results show that of the samples from Asia only 3% were from the Ursidae family and 18% were from "farmed bears." Samples seized in the U.S.A. and Canada showed that 22.6% and 85% respectively, were from Ursids. The remaining samples were consistent with bile from the domestic pig (Suidae).
Assuntos
Ácidos e Sais Biliares/análise , Vesícula Biliar/química , Ursidae/classificação , Animais , Ásia , Comércio , Medicina Legal , América do NorteRESUMO
The biliary bile acid composition of gallbladder bile obtained from six species of bears (Ursidae), the Giant panda, the Red panda, and 11 related carnivores were determined by reversed phase liquid chromatography and gas chromatography-mass spectrometry. Bile acids were conjugated solely with taurine (in N-acyl linkage) in all species. Ursodeoxycholic acid (3 alpha, 7 beta-dihydroxy-5 beta-cholan-24-oic acid) was present in all Ursidae, averaging 1-39% of biliary bile acids depending on the species; it was not detected or present as a trace constituent (< 0.5%) in all other species, including the Giant panda. Ursodeoxycholic acid was present in 73 of 75 American Black bears, and its proportion averaged 34% (range 0-62%). Ursodeoxycholic acid averaged 17% of biliary bile acids in the Polar bear (n = 4) and 18% in the Brown bear (n = 6). Lower proportions (1-8%) were present in the Sun bear (n = 2), Ceylon Sloth bear (n = 1), and the Spectacled bear (n = 1). Bile of all species contained taurine-conjugated chenodeoxycholic acid and cholic acid. In some related carnivores, deoxycholic acid, the 7-dehydroxylation product of cholic acid, was also present. To determine whether the 7 beta hydroxy group of ursodeoxycholic acid was formed by hepatic or bacterial enzymes, bile acids were determined in hepatic bile obtained from bears with chronic biliary fistulae. Fistula bile samples contained ursodeoxycholic acid, chenodeoxycholic acid, and a trace amount of cholic acid, all as taurine conjugates, indicating that ursodeoxycholic acid is a primary bile acid formed in the liver in Ursidae.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Ácidos e Sais Biliares/análise , Bile/química , Carnívoros/metabolismo , Ursidae/metabolismo , Ácido Ursodesoxicólico/análise , Animais , Animais Recém-Nascidos , Ácido Quenodesoxicólico/análise , Ácido Cólico , Ácidos Cólicos/análise , Cromatografia Líquida de Alta Pressão , Ácido Desoxicólico/análise , Valores de Referência , Especificidade da Espécie , Taurina/metabolismoRESUMO
The role of bile acids in the regulation of cholesterol 7 alpha-monooxygenase (EC 1.14.13.17) was characterized using primary cultures of rat hepatocytes supplemented with dexamethasone and thyroxine. Taurocholate and taurodeoxycholate (50 microM) repressed cholesterol 7 alpha-hydroxylase mRNA to 44 +/- 9 and 52 +/- 4%, respectively, of control values. Repression by these natural, relatively hydrophobic bile acids was concentration dependent, with an IC50 of about 50 microM, and time dependent with a t1/2 for repression of 22 h. In contrast, two natural hydrophilic bile acids, tauroursodeoxycholate and taurohyodeoxycholate, had no effect on cholesterol 7 alpha-hydroxylase mRNA levels. Taurochenodeoxycholate and taurolithocholate also had no effect, but these hydrophobic bile acids were rapidly hydroxylated to more hydrophilic bile acids. Hydrophilic bile acid analogues (nor (C23) bile acids and beta-hydroxy epimers) repressed cholesterol 7 alpha-hydroxylase mRNA less potently than their corresponding and more hydrophobic C24 or alpha-hydroxy derivatives. Cholesterol 7 alpha-hydroxylase specific activity was decreased by taurocholate or taurodeoxycholate (50 microM) to 26 +/- 9 and 56 +/- 3% of control, respectively; its transcriptional activity was repressed to 52 +/- 5% of control by taurocholate (50 microM). The addition of cholesterol or the induction of cholesterol biosynthesis did not influence repression of cholesterol 7 alpha-hydroxylase mRNA levels by taurocholate. Based on several lines of evidence, cAMP was not involved in bile acid-induced repression. In rat hepatocytes cultured under conditions in which cholesterol 7 alpha-hydroxylase gene expression is maintained at in vivo levels, hydrophobic bile acids repress this enzyme at the level of gene transcription independently of cholesterol availability.