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1.
Human hepatic beta-glucuronidase: an enzyme kinetic study   总被引:1,自引:0,他引:1  
Y C Ho  L H Ho  K J Ho 《Enzyme》1985,33(1):9-17
beta-Glucuronidase (EC 3.2.1.31) was purified from human liver and its activity was determined by enzyme kinetic method employing phenolphthalein glucuronic acid (PGA) and conjugated bilirubin, primarily bilirubin diglucuronide purified from human bile, as substrates in the absence or presence of D-glucaro-1,4-lactone. The enzyme was capable of acting on both PGA and conjugated bilirubin with Michaelis constants of 0.435 mmol/l at 56 degrees C and 1.02 mmol/l at 37 degrees C, respectively. Both reactions were beta-glucuronidase-specific because both were inhibited by D-glucaro-1,4-lactone in a competitive fashion. Conjugated bilirubin acted as a noncompetitive inhibitor of the enzyme for PGA in a two-substrate system. The study indicates that these two substrates bind at different catalytic sites of the enzyme and, on molar base, conjugated bilirubin had higher affinity for the enzyme and less degree of inhibition by D-glucaro-1,4-lactone than PGA. Whether such catalytic sites are also common for other beta-D-glucuronid ethers and esters remains to be proven.  相似文献   

2.
The net glucuronidation of bilirubin (BR) has been determined in inbred and outbred rat strains and their subpopulations with similar glucuronosyltransferase (EC 2.4.1.17) activity but with different levels of beta-glucuronidase (beta G) (EC 3.2.1.31), or in which the level of beta G activity was reduced with D-glucaro-1,4-lactone. These studies demonstrated that outbred rat strains consist of two subpopulations that differ approximately 1.5- to two-fold in serum and liver beta G activity. Evidence is presented indicating that owing to its compartmentalization the lysosomal beta G, unlike the corresponding microsomal enzyme, is neither inhibited by glucarolactone nor accessible for hydrolysis of newly synthesized glucuronides. The ratio of glucuronidated to unconjugated BR 15 min after injection of albumin-bound BR into the tail vein appears to correlate negatively with the liver microsomal beta G activity. The results may be relevant to the relative risk to toxins, including carcinogens, and to their reduction by dietary intervention.  相似文献   

3.
Quercetin glucuronides are the main circulating metabolites of quercetin in humans. We hypothesise that the potential availability of the aglycone within tissues depends on the substrate specificity of the deconjugating enzyme beta-glucuronidase towards circulating flavonoid glucuronides. Human tissues (small intestine, liver and neutrophils) exhibited beta-glucuronidase against quercetin glucuronides. The various quercetin glucuronides were deconjugated at similar rates, but liver cell-free extracts were the most efficient and the activity was completely inhibited by saccharo-1,4-lactone (a beta-glucuronidase inhibitor). Furthermore, pure recombinant human beta-glucuronidase hydrolysed various flavonoid glucuronides, with a 20-fold variation in catalytic efficiency (k(cat)/K(m)=1.3x10(3) M(-1) s(-1) for equol-7-O-glucuronide and 26x10(3) M(-1) s(-1) for kaempferol-3-O-glucuronide). Similar catalytic efficiencies were obtained for quercetin O-glucuronides substituted at different positions. These results show that flavonoid glucuronides can be deconjugated by microsomal beta-glucuronidase from various human cells.  相似文献   

4.
To investigate the modifying role of the intestinal microflora in the metabolism of 1-nitropyrene (1-NP) via enterohepatic circulation, we collected bile from male Wistar rats administered [3H]1-NP orally. The bile was mixed with the intestinal contents (IC) prepared from untreated rats and the mixture was incubated anaerobically under an atmosphere of nitrogen at 37 C. Samples of the reaction mixture were removed at intervals to assay their mutagenic potential, to determine the radioactivity bound to the IC, and for analysis of the biliary metabolites. The binding of the radioactivity to the IC increased linearly as a function of time during the 1-hr incubation. The time-dependent binding does not occur with heat-treated IC and the binding was inhibited by addition of D-saccharic acid 1,4-lacton, a beta-glucuronidase inhibitor. The mutagenicity (for Salmonella typhimurium strain TA98 without S9 mix) of the bile increased early in the incubation period and then decreased very rapidly. The mutagenicity of the bile was also enhanced by treatment with a sonicated IC extract or beta-glucuronidase, but not with a heat-treated IC or aryl-sulfatase. The metabolites produced after the bile was incubated for short periods with the IC were mainly nitrohydroxypyrenes; at later times nitroreduction occurred. The level of acetylaminohydroxypyrenes, which were formed by deconjugation, did not change during the incubation. To determine the degree of contribution of the IC to the total acetylating capacity, we measured acetyltransferase activity of the IC and various organs in Wistar rats. The liver had the highest N-acetyltransferase activity among the seventeen organs examined. Considerable activity was also detected in the kidney, small intestine, lung, and testis, but the IC showed very low activity. The acetylating capacity of the IC was 0.27% of the total capacity in rats, and that of the liver was more than 80%. These results suggest that the nitrohydroxypyrenes formed from 1-NP in the liver were conjugated to glucuronic acid and excreted via the bile duct into intestine. Hydrolysis of these glucuronide conjugates by bacterial beta-glucuronidase liberated into intestine, free nitrohydroxypyrenes, which were direct-acting mutagens. The released aglycons were then rapidly nitro-reduced by intestinal microflora, but contribution of the intestinal microflora to acetylation of the reduced metabolites is very low.  相似文献   

5.
At high dietary levels in vivo, both 13-cis-retinoic acid and calcium glucarate inhibit the induction of rat mammary tumors by 7,12-dimethylbenz(a)anthracene. The present study shows that sub-optimal dietary levels of each, which individually have no effect on tumor induction, when combined together in the diet, significantly increases tumor latency and suppresses tumor frequency in the rat system. Weight gain of animals was similar in control and experimental groups. Furthermore, ineffective sub-optimal dosages of glucarate and 13-cis-retinoic acid interacted synergistically to inhibit the growth in vitro of the MCF-7 human breast cancer cells. By varying the concentrations of glucarate and 13-cis-retinoic acid independently, evidence was obtained that in combination glucarate may play an adjuvant role, with the retinoid as the effector. Thus, the results of this experimental animal study demonstrate for the first time the potential use in synergistic combination of 2 normal metabolites in non-toxic chemoprevention and chemotherapy.  相似文献   

6.
Glucarate hydro-lyase was isolated and purified to near homogeneity from cells of Pseudomonas acidovorans grown on glucarate. By using gel filtration and ion-exchange chromatography, it was shown that the oxodeoxyglucarate aldolase activity observed in such extracts is associated with the glucarate hydro-lyase protein.  相似文献   

7.
1. A radiochemical method for the studies on the microsomal UDPglucuronic acid metabolism has been developed. 2. The rat liver microsomes caused a rapid hydrolysis of UDPglucuronic acid to D-glucuronic acid 1-phosphate and further although much slower to free D-glucuronic acid. In Tris-HCl buffer (pH 7.4) they were produced in ratio 72 : 1. No other metabolites were found in measurable amounts. The pyrophosphatase splitting UDPglucuronic acid showed a pH optimum at 8.9, but the liberation of D-glucuronic acid from UDPglucuronic acid had two pH maxima (pH 3.5 and 8.5). EDTA appeared to be less powerful inhibitor of pyrophosphatase than previously suggested. About 25 per cent of the UDPglucuronic acid hydrolyzing activity was still remaining in the presence of 10 mM EDTA. D-Glucaro-1,4-lactone was found to have a slight inhibitory action on the pyrophosphatase activity. Citrate inhibited powerfully the hydrolysis of UDPglucuronic acid and the liberation of free D-glucuronic acid. Phosphate was also inhibitory. 3. In the presence of an exogenous UDPglucuronosyltransferase substrate, 4-nitrophenol, the formation of D-glucuronic acid 1-phosphate and free D-glucuronic acid were slightly reduced, and D-glucuronic acid 1-phosphate, 4-nitrophenylglucuronide and free D-glucuronic acid were produced in ratio 78 : 23 : 1. When 10 mM EDTA was added to diminish the hydrolytic consumption of the glucuronyl donor substrate, the corresponding ratio was still as unfavorable as 19 : 2.6 : 1. The measurable activity of UDPglucuronosyltransferase was lower in the presence of phosphate or citrate than in Tris-HCl buffer, although they protected the glucuronyl donor substrate against hydrolysis. 4. The results indicate that even in the presence of added glucuronyl acceptor substrate the hydrolysis of UDPglucuronic acid predominates the conjugation in rat liver microsomes. The rate of the hydrolysis of UDPglucuronic acid is quite considerable even in the presence of EDTA, and it is recommended to control the UDPglucuronic acid pyrophosphatase activity when UDPglucuronosyltransferase and glucuronidation reactions are studied. Free D-glucuronic acid appears to be produced from UDPglucuronic acid for further use via D-glucuronic acid 1-phosphate, the rate-limiting step being the hydrolysis of this intermediate. UDP-glucuronosyltransferase, glucuronides of either endogenous or exogenous aglycones and beta-glucuronidase have only a minor role in this respect in rat liver microsomes.  相似文献   

8.
The glycoprotein egasyn complexes with and stabilizes precursor beta-glucuronidase in microsomes of several mouse organs. Several observations indicate egasyn is, in addition, an esterase. Liver homogenates of egasyn-positive strains have specific electrophoretically separable esterases which are absent in egasyn-negative mice. These esterases react with anti-egasyn serum. A specific esterase was likewise complexed with immunopurified microsomal beta-glucuronidase. The esterases were, like egasyn and microsomal beta-glucuronidase, concentrated in the microsomal subcellular fraction. Egasyn which is not bound to beta-glucuronidase, which represents 80-90% of total liver egasyn, is not complexed with other liver proteins. Egasyn, therefore, specifically stabilizes beta-glucuronidase in microsomes. The esterase activity is inhibited by bis-p-nitrophenyl phosphate indicating it is a carboxyl esterase. Several possible functions of egasyn-esterase activity are discussed.  相似文献   

9.
Aldehydes released during peroxidation of liver microsomal lipids and identified as 4-hydroxyalkenals (4-hydroxynonenal being quantitatively the most significant) strongly inhibited the calcium sequestration activity of liver microsomes. The ID50 for 4-hydroxynonenal was 42 microM. The inhibition appeared to be correlated with the amount of the aldehyde bound to the microsomal protein. In rats intoxicated with BrCCl3 significant amounts of protein-bound aldehydes were formed at only 5 min after poisoning, a time at which the calcium sequestring capacity is markedly inhibited.  相似文献   

10.
The regulation of hepatic and intestinal 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase and acyl-CoA; cholesterol acyltransferase (ACAT) activities by dietary fish oil was examined in the rabbit. Rabbits were fed 10% menhaden oil or menhaden oil plus 1% cholesterol for 14 days. They were compared with animals fed a control diet or one enriched with long-chain saturated fats consisting of 10% cocoa butter oil or cocoa butter oil plus 1% cholesterol. Plasma cholesterol was increased in rabbits fed the fish oil and the two cholesterol-containing diets. In the liver, ACAT activity was increased and HMG-CoA reductase activity was decreased in rabbits ingesting the fish oil. The same was true for animals ingesting both cholesterol-containing diets. In the intestine, ACAT activity was not affected by the ingestion of the fish oil compared to control rabbits; however, it was significantly higher in animals fed the fish oil compared to animals ingesting the cocoa butter. HMG-CoA reductase activity was decreased in the distal two-thirds of the intestine in animals fed the menhaden oil compared to activities observed in controls. In animals ingesting the cholesterol diets, intestinal reductase was significantly decreased, whereas intestinal ACAT activity was increased in rabbits ingesting the cocoa butter and cholesterol diet when compared to their controls. Lipid analysis of hepatic and intestinal microsomes demonstrated an enrichment of n-3 polyunsaturated fatty acids in membranes from rabbits ingesting the menhaden oil.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
Heme oxygenase activity was examined in the epithelial cells of the small intestine in male Sprague-Dawley rats. As with liver and spleen, the highest specific activity of this enzyme was found in the microsomal fraction of these cells. Substrate kinetics, analysis of cofactor requirements, and other biochemical characteristics suggested further similarities between heme oxygenase in the small intestine and liver. Enzyme activity was differentially localized longitudinally within the small intestine, with the highest specific activity occurring in the region approximately 15 to 30 cm beyond the pylorus. The effects of diet on the basal levels of heme oxygenase in the proximal small intestine were also examined. Although intestinal cytochrome P450-dependent monooxygenase activity, as determined by benzo[a]pyrene hydroxylase and 7-ethoxycoumarin O-deethylase, was greatly reduced (65-90%) in animals maintained on a semipurified control diet compared with standard cereal-based chow, there were no differences observed in heme oxygenase activity between the two dietary treatment groups. The activity of intestinal heme oxygenase could be increased, however, by oral treatment with several metal compounds that are known to affect hepatic heme metabolism when administered parenterally. The enzyme activity was also potently inhibited by tin (Sn4+) protoporphyrin administered orally or parenterally.  相似文献   

12.
The mechanisms by which the in vivo intoxication with BrCCl3 inhibits the calcium sequestration activity of liver microsomes were studied. The initial rate of Ca2+ transport is inhibited by nearly 50% in the intoxicated rats as compared to the controls; this indicates that the active transport of Ca2+ is markedly affected by the intoxication. The microsomal ATPases activities both in the presence and in the absence of Ca2+ were not decreased at all in the intoxicated animals. However, the Ca2+-dependent extra ATP hydrolysis shows a different kinetics in the BrCCl3-poisoned rats with respect to the controls. The release of Ca2+ from Ca2+ loaded liver microsomes is higher in the intoxicated animals. It seems therefore that the increased permeability of the membrane to Ca2+ contributes to some extent to the haloalkane-induced inhibition of the calcium sequestration activity of liver microsomes.  相似文献   

13.
Ethylmorphine N-demethylase activity of the sheep liver and lung microsomes was reconstituted in the presence of solubilized microsomal cytochrome P-450, NADPH-cytochrome c reductase and synthetic lipid, phosphatidylcholine dilauroyl. The Km of the lung microsomal ethylmorphine N-demethylase was calculated to be 4.84 mM ethylmorphine from its Lineweaver-Burk graph and lung enzyme was inhibited by its substrate, ethylmorphine, when its concn was 25 mM and above, reaching to 67% inhibition at 50 mM concn. The Lineweaver-Burk and Eadie-Hofstee plots of the liver enzyme were found to be curvilinear. From these graphs, two different Km values were calculated for the liver enzyme as 4.17 mM and 0.40 mM ethylmorphine. Ethylmorphine N-demethylase activities of both liver and lung microsomes were inhibited by NiCl2, CdCl2 and ZnSO4. Ethylalcohol inhibited N-demethylation of ethylmorphine in lung and liver microsomes. Acetone (5%) slightly enhanced the N-demethylase activity of the liver enzyme, whereas 5% acetone completely inhibited the lung enzyme. Phenylmethylsulfonyl fluoride at 0.10 mM and 0.25 mM concn had no effect on liver enzyme activity, while at these concns, it inhibited the activity of the lung enzyme by about 35%.  相似文献   

14.
Treatment of 2-acetamido-2-deoxy-D-mannono-1,4-lactone with dicyclohexylamine in ethanolic solution afforded an unsaturated 1,4-lactone, 2-acetamido-2,3-dideoxy-D-erythro-hex-2-enono-1,4-lactone (1), in good yield. 2-Acetamido-2,3-dideoxy-D-threo-hex-2-enono-1,4-lactone (2) was similarly prepared from 2-acetamido-2-deoxy-D-galactono-1,4-lactone. An unsaturated 1,5-lactone, 2-acetamido-2,3-dideoxy-D-threo-hex-2-enono-1,5-lactone (4), was obtained through the oxidation of 2-acetamido-2-doexy-4,6-0-isopropylidene-D-galactopyranose with silver carbonate on Celite, followed by mild hydrolysis. The inhibitory activity of four isomeric 2-acetamido-2,3-dideoxy-D-hex-2-enonolactones [1, 2, 4, and 2-acetamido-2,3-dideoxy-D-erythro-hex-2-enono-1,5-lactone (3)] was assayed against 2-acetamido-2-deoxy-beta-D-glucosidase from bull epididymis. Only the erythro lactones 1 and 3 are weak competitive inhibitors, whereas the threo lactones 2 and 4 are practically inactive. The 1,4-lactone 1 inhibited 2-acetamido-2-deoxy-beta-D-glucosidase more strongly than the 1,5-lactone 3. The lactones 1-4 were found to be quite stable in aqueous solution or under inhibitory-assay conditions. In addition, two 2-acetamido-2-deoxy-D-glycals, 2-acetamido-1,5-anhydrohex-1-enitol (7) were tested; both are 10 times as active as 1.  相似文献   

15.
Citroylformic acid-γ-lactone (CFA, 1-keto-2,4-dihydroxy-4-carboxyadipenoic acid(2–3)-1,4-lactone), isolated from a commercial batch of oxaloacetate, inhibited conversion of rat liver apotyrosine aminotransferase (EC 2.6.1.5) to holoenzyme. Using partially purified enzyme, the Ki was determined to be less than 0.7 mm. A more definitive Ki was difficult to obtain because at pH 7 CFA had a half-life of about 2 hr. Inhibition of the enzyme by CFA was stereospecific and reversible; the S (?) stereoisomer was approximately 10 times more inhibitory than its R(+) antipode, and over 90% of inhibited enzyme was recoverable after overnight dialysis. Preineubation of apotyrosine aminotransferase with its coenzyme (pyridoxal phosphate) prevented inhibition by CFA, and a substantial fraction of enzyme that had been inhibited by CFA could be readily reactivated by addition of high concentrations of pyridoxal phosphate. Studies with inhibitor analogs indicated that both a partially unsaturated lactone ring and a stereospecific carboxymethyl group are required for maximal inhibitory activity. The sodium salts of citroylformic acid and oxalopyruvic acid, formed by the hydrolysis of their respective lactones, were not inhibitory; 1-keto-2,4-dihydroxy-4-carboxyadipic acid-γ-lactone and little inhibitory activity, and 1-keto-2,4-dihydroxyglutarenoic acid-γ-lactone and 1-keto-2,4-dihydroxybutene-γ-lactone were somewhat better inhibitors than the R(+) stereoisomer of CFA. The possibility that CFA is a naturally occurring biological substance is discussed.  相似文献   

16.
Hukmani P  Tripathy BC 《Plant physiology》1994,105(4):1295-1300
The inhibitor sensitivity of the endoplasmic reticulum (ER) and plasma membrane (PM) calcium pumps of red beet (Beta vulgaris L.) were studied by measuring the ATP-driven accumulation of 45Ca2+ into isolated membrane vesicles. Both transporters were strongly inhibited by 50 [mu]mol m-3 erythrosin B, but only by 50% in the presence of 100 mmol m-3 vanadate. A number of inhibitors considered to be specific for the sarcoplasmic reticulum (SR)/ER-type calcium pump in animal cells were used to further characterize the PM and ER Ca2+-ATPases in red beet and were compared with their effect on the transport and hydrolytic activities of the PM and tonoplast H+-ATPases. The hydroquinones 2,5-di(tert-butyl)-1,4-benzohydroquinone and 2,5-di(tert-amyl)-1,4-benzohydroquinone produced around 20 and 40% inhibition of activity, respectively, of the PM and ER calcium pumps and the PM H+-ATPase when present at concentrations of 30 mmol m-3. In contrast, the vacuolar proton pump displayed a much higher sensitivity to these two compounds. Nonylphenol appeared to have a general inhibitory effect on all four membrane transport proteins and gave almost complete inhibition when present at a concentration of 100 mmol m-3. Thapsigargin and the structurally related compound trilobolide produced 50% inhibition of both the ER and PM calcium pumps at concentrations of 12.5 and 24 mmol m-3, respectively. The PM and tonoplast proton pumps were also sensitive to these compounds. The ER and PM calcium pumps were almost completely insensitive to cyclopiazonic acid (CPA) up to a concentration of 20 mmol m-3. When present at 100 mmol m-3 CPA caused 30% inhibition of the transport properties of all four ATPases. The high concentrations of all of the inhibitors of the SR/ER Ca-ATPase required to inhibit the red beet ER calcium pump, together with the similar effects on the PM calcium pump and the PM and tonoplast proton pumps, suggests that these hydrophobic compounds have a general nonselective action in red beet, possibly through disruption of membrane lipid-protein interactions.  相似文献   

17.
The subcellular localization of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in rat intestine was reinvestigated. Highly enriched fractions of endoplasmic reticulum and mitochondria were prepared from mucosal cells. The highest specific activity of HMG-CoA reductase was located in the endoplasmic reticulum fraction with recovery of 25% of the total activity. The mitochondria had low specific activity and low recovery of reductase activity relative to whole homogenate (2-5%). Despite attempts to maximize cell lysis, much of the activity (about 60%) was recovered in a low speed pellet which consisted of whole cells, nuclei, and cell debris as determined by light microscopy. Taken together, the evidence strongly suggests that much of the cellular HMG-CoA reductase activity is present in the endoplasmic reticulum fraction and that mitochondria have little or no intrinsic HMG-CoA reductase. The in vitro regulation of intestinal microsomal HMG-CoA reductase was studied. The intestine possesses a cytosolic HMG-CoA reductase kinase-phosphatase system which appears to be closely related to that present in the liver. Intestinal reductase activity in microsomes prepared from whole mucosal scrapings was inhibited 40-50% by the presence of 50 mM NaF in the homogenizing buffer. It was less susceptible to the action of the kinase than liver reductase. The effects of NaF were reversed by incubation with partially purified intestinal or liver phosphatases. These results suggest that the kinase-phosphatase system could play a role in the regulation of intestinal sterol and isoprene synthesis in vivo.  相似文献   

18.
Forty-eight adult hamsters were divided equally into two groups fed a control diet and a 2% cholesterol diet, respectively, under a rigid lighting (6 PM-6 AM) and feeding (6 PM-8 AM) schedule for three weeks. The cholesterol synthetic activity of the liver, stomach, small intestine, cecum, colon and kidney was measured by in vivo conversion of acetate-1-14C to cholesterol in four animals each time at 4 hour intervals. A remarkable circadian rhythm with the peak at midnight and the nadir at noon was found in the liver of the control hamsters, but was completely abolished in the cholesterol-fed animals since the activity was nearly totally suppressed at all times. The small intestine exhibited a similar rhythm with the peak at midnight but maintained a high baseline activity from 8 AM to 6 PM. Cholesterol feeding did not alter the baseline activity but reduced 17% of the peak activity. Other organs failed to show such a circadian rhythm.  相似文献   

19.
We determined the site of synthesis of apolipoprotein (apo) E and apo-A-I in rabbit by measuring in vitro translational activity of their mRNAs from the liver and from the intestine. Poly(A+) RNA isolated from liver and intestinal epithelium of rabbits fed either a chow diet or a cholesterol-rich diet was translated in vitro in the rabbit reticulocyte lysate system using [35S] methionine as the labeled precursor. Newly synthesized apolipoproteins were immunoprecipitated with specific antisera and quantitated after electrophoresed on 10% polyacrylamide slab gels in the presence of 0.2% sodium dodecyl sulfate. The levels of liver apo-E and apo-A-I mRNAs from chow-fed rabbits are 0.41 and 0.002% of total translatable mRNA, respectively. The level of liver apo-A-I mRNA in the rabbit is approximately 500-fold lower than the reported level of apo-A-I mRNA in rat and human livers. Rabbit intestinal apo-E and apo-A-I mRNAs levels are 0.0036 and 0.67%, respectively. Our results indicate that in rabbits apo-E is synthesized primarily in the liver and that apo-A-I is synthesized primarily in the intestine. When rabbits are fed a cholesterol-rich diet, liver and intestinal apo-E in mRNA levels and intestinal apo-A-I mRNA levels are not changed. In contrast, the liver apo-A-I mRNA level increases 5-fold in response to the cholesterol-rich diet. However, because the intestinal liver apo-A-I mRNA level is so low, the 5-fold induction only increases liver mRNA levels to 2.7% of the corresponding intestinal apo-A-I mRNA level.  相似文献   

20.
Rates of production of p-nitrophenyl glucuronide by isolated perfused livers from fed or fasted phenobarbital-treated rats were estimated by monitoring the concentration of glucuronide in the effluent perfusate. Infusion of epinephrine decreased the steady state level of p-nitrophenyl glucuronide by about 39% (half-maximal inhibition at approximately 5 microM). This result was unexpected because epinephrine activated glycogenolysis and elevated hepatic UDP-glucuronic acid contents. The effect of epinephrine can be attributed to its interaction with alpha-adrenergic receptors, since the inhibition of glucuronide production by epinephrine was reversed by an alpha-antagonist (phentolamine) but not by a beta-antagonist, propranolol. Since alpha-adrenergic agonists increase the cytosolic free calcium concentration, we investigated the possibility that the decrease in glucuronide production elicited by epinephrine was mediated by calcium. Removal of calcium from the perfusion fluid diminished the inhibition of glucuronide production by epinephrine, while increasing extracellular calcium from 0 to 150 microM restored the inhibition in a dose-dependent manner. In the presence of extracellular calcium, glucuronide production was inhibited by the addition of the calcium ionophore A23187 or angiotensin II, a hormone which increases cytosolic calcium. Concentrations of ionized calcium comparable to physiological intracellular levels (0.1-2 microM) increased microsomal beta-glucuronidase activity by 50 to 100% but had no effect on microsomal glucuronosyl-transferases . These results indicate that activation of hepatic alpha-adrenergic receptors increases cytosolic calcium which stimulates microsomal beta-glucuronidase activity. This decreases net glucuronide formation by the liver. In support of this hypothesis, rates of glucuronide production were unaffected by epinephrine in perfused livers from beta-glucuronidase-deficient C3H/HeJ mice.  相似文献   

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