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1.
Metabolic fate of endogenously synthesized prostaglandin D2 in a human female with mastocytosis 总被引:4,自引:0,他引:4
Increased production of prostaglandin D2 was recently demonstrated in patients with systemic mastocytosis. One female patient investigated with mastocytosis was found to have overproduction of prostaglandin D2 of such magnitude (150-fold above normal) that it provided the unique opportunity to delineate the metabolic fate of endogenously synthesized prostaglandin D2. A five percent aliquot of a twenty-four hour urine collection from this patient was extracted, purified by silicic acid chromatography, methylated, and finally subjected to high pressure liquid chromatography. Column fractions collected were derivatized and analyzed by gas chromatography-mass spectrometry. Increased quantities of sixteen urinary metabolites were identified and included a series of metabolites retaining the PGD-ring as well as a series of metabolites with a PGF-ring. PGF-ring metabolites were excreted in approximately 4-fold greater relative abundance than PGD-ring metabolites. More than one apparent isomeric form of some PGF-ring metabolites were found. The predominant urinary metabolite was 2,3-dinor-prostaglandin F2. This study provides evidence that endogenously synthesized prostaglandin D2 is converted in substantial part to prostaglandin F2 metabolites in vivo in humans. 相似文献
2.
Joseph H. Butterfield 《Prostaglandins & other lipid mediators》2009,88(3-4):122-124
BackgroundPrevious recommended doses for aspirin use in systemic mastocytosis have been 3.9–5.2 g/d. Here, the aspirin doses and biochemical responses of patients with systemic mastocytosis given aspirin to decrease prostaglandin D2 levels and prevent symptoms were reviewed.MethodsTwenty patients with systemic mastocytosis who had been given aspirin were identified, and their clinical and laboratory records were reviewed including changes in the excretion of the prostaglandin D2 metabolite 11β-prostaglandin F2α in response to aspirin.ResultsTwo of 20 patients developed either a delayed reaction or flushing during outpatient updosing with aspirin. In 20 of 20 patients with elevated baseline urinary excretion of 11β-prostaglandin F2α, aspirin therapy caused a reduction to normal levels of excretion. Doses of aspirin required ranged from 81 mg twice daily to 500 mg twice daily.ConclusionsControl of elevated prostaglandin D2 levels in systemic mastocytosis can be achieved with lower doses of aspirin than previously reported as necessary in this disorder. 相似文献
3.
Tsuyoshi Watanabe Takao Shimizu Shuh Narumiya Osamu Hayaishi 《Archives of biochemistry and biophysics》1982,216(1):372-379
Two forms of NADP-linked 15-hydroxyprostaglandin dehydrogenase for prostaglandin D2 were found in the cytosol fraction of human blood platelets. These enzymes were purified by ammonium sulfate fractionation, Blue Sepharose, and Sephadex G-100 column chromatography. The two enzymes differed in molecular weights (65,000 for peak I enzyme and 31,000 for peak II as estimated by gel filtration) and their substrate specificities. The relative rates for reaction with peak I enzyme were: prostaglandin D2, 100(%); E2, 14; F2α, 2; I2, 29; and B2, 0; whereas for peak II enzyme, D2, 100; E2, 23; F2α, 61; I2, 29; and B2, 131. Prostaglandin D2 was converted to 15-ketoprostaglandin D2 and then 13,14-dihydro-15-ketoprostaglandin D2, which were identified by spectrophotometry and gas chromatography/mass spectrometry, respectively. These metabolites were three orders of magnitude less potent in inhibiting human platelet aggregation than prostaglandin D2. The results indicated that NADP-linked dehydrogenases participated in the metabolic inactivation of prostaglandin D2 in the platelets. Furthermore, the dehydrogenase activity for prostaglandin D2 was high in monkey (0.128 nmol/min · mg at 24 °C) and human platelets (0.066), but was not detectable (less than 0.007) in the rabbit, rat, and chicken. Because prostaglandin D2, which was demonstrated by several authors to be synthesized in platelet-rich plasma during platelet aggregation, exhibited significant antiaggregatory activity only in human and monkey platelets, these prostaglandin dehydrogenases appear to play a physiological role in the circulatory system. 相似文献
4.
50 microCi of [3H]prostaglandin D2 tracer (100 Ci/mmol) was infused intravenously into a normal human male volunteer. 75% of the infused radioactivity was excreted into the urine within 5 h. This urine was added to urine obtained from two mastocytosis patients with marked overproduction of prostaglandin D2. Radiolabeled prostaglandin D2 urinary metabolites were chromatographically isolated and purified and subsequently identified by gas chromatography-mass spectrometry. 25 metabolites were identified. 23 of these compounds comprising 37% of the recovered radioactivity had prostaglandin F-ring structures, and only two metabolites comprising 2.7% of the recovered radioactivity retained the prostaglandin D-ring structure. The single most abundant metabolite identified was 9,11-dihydroxy-15-oxo-2,3,18,19-tetranorprost-5-ene-1,20-dioic acid which was isolated in a tricyclic form as a result of formation of a lower side chain hemiketal followed by lactonization of the terminal carboxyl and the hemiketal hydroxyl. Different isomeric forms of several prostaglandin F-ring metabolites were identified. An isomer of prostaglandin F2 alpha was also excreted intact into the urine as a metabolite of prostaglandin D2. 15 PGF-ring compounds were treated with n-butylboronic acid and 13 failed to form a boronate derivative, suggesting that the orientation of the hydroxyl group at C-11 in these 13 metabolites is beta. This study documents that prostaglandin D2 is metabolized to prostaglandin F-ring metabolites in vivo in humans. These results also bring into question the accuracy of quantifying prostaglandin F2 alpha metabolites as a specific index of endogenous prostaglandin F2 alpha biosynthesis, as well as quantifying urinary prostaglandin F2 alpha as an accurate index of renal production of prostaglandin F2 alpha. 相似文献
5.
《Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism》1987,917(3):356-364
The metabolic transformation of exogenous prostaglandin D2 was investigated in isolated perfused rat lung. Dose-dependent formation (2–150 ng) of 9α,11β-prostaglandin F2, corresponding to about 0.1% of the perfused dose of prostaglandinD2, was observed by specific radioimmunoassay both in the perfusate and in lung tissue after a 5-min perfusion. To investigate the reason for this low conversion ratio, we analyzed the metabolites of tritium-labeled 9α,11β-prostaglandin F2 and prostaglandin D2 by boric acid-impregnated TLC and HPLC. By 5 min after the start of perfusion, 9α,11β-prostaglandin F2 disappeared completely from the perfusate and the major product formed remained unchanged during the remainder of the 30-min perfusion. The major product was separated by TLC and identified as 13,14-dihydro-15-keto-9α,11β-prostaglandin F2 by GC/MS. In contrast, pulmonary breakdown of prostaglandin D2 was slow and two major metabolites in the perfusate increased with time, each representing 56% and 11% of the total radioactivity at the end of the perfusion. The major product (56%) was identified as 13,14-dihydro-15-ketoprostaglandin D2 and the minor one (11%) was tentatively identified as 13,14-dihydro-15-keto-9α,11β-prostaglandin F2 based on the results from radioimmunoassays, TLC, HPLC, and the time course of pulmonary breakdown. These results demonstrate that the metabolism of prostaglandin D2 in rat lung involves at least two pathways, one by 15-hydroxyprostaglandin dehydrogenase and the other by 11-ketoreductase, and that the 9α,11β-prostaglandin F2 formed is rapidly metabolized to 13,14-dihydro-15-keto-9α,11β-prostaglandin F2. 相似文献
6.
. Gecse Anna Ottlecz Zsfia Mezei G. Telegdy F. Jo E. Dux Irina Karnushina 《Prostaglandins & other lipid mediators》1982,23(3):287-297
The synthesis of prostacyclin and prostaglandins was examined in isolated blood-free brain capillaries of guinea-pigs and rats using 1-14C-arachidonic acid as a precursor. The main prostaglandins synthesized by guinea-pig microvessels were prostaglandin D2 and prostaglandin E2. Substantially less prostaglandin F2α or the prostacyclin stable metabolite, 6-oxo-prostaglandin F1α was synthesized. Rat capillary prostaglandin distribution differed substantially from that of the guinea-pigs although the principle prostaglandin was also PGD2. Total prostaglandin conversion was greater in guinea-pig capillaries than in the rat.Norepinephrine stimulated the prostaglandin forming capacity of blood free cerebral microvasculature of guinea-pigs. Prostacyclin and prostaglandins could be involved in the activity dependent regulation of regional cerebral blood flow and permeability. 相似文献
7.
A highly sensitive and specific radioimmunoassay for prostaglandin D2 has been developed and used to determine the basal level and regional distribution of this prostaglandin in rat brain, spinal cord and pituitary. The assay can detect as little as 20 pg of prostaglandin D2, and the antiserum used shows 20% cross-reactivity to prostaglandin D1, 0.1% cross-reactivity to 13,14-dihydro-15-ketoprostaglandin D2 and even lower cross-reactivity to other prostaglandins. Prostaglandin D2-like immunoreactivity was extracted with ethanol from the rat tissues. The immunoreactivity comigrated with authentic prostaglandin D2 on silica gel thin layer chromatography, showed the dilution curve parallel to that of the authentic compound, and decreased in amounts by the pretreatment of animals with indomethacin, suggesting that it was prostaglandin D2 itself. To avoid a postmortem formation of prostaglandins, we sacrificed animals by microwave irradiation at 4.5 kW for 1.2 sec under which conditions both prostaglandin D synthetase and prostaglandin D dehydrogenase were completely inactivated. The amount of prostaglandin D2 in whole brain measured under these conditions was 3.42±0.59 ng (mean+S.E.M.), and those of prostaglandin E2 and F2α measured by the respective radioimmunoassays were 1.32±0.24 and 0.96±0.20 ng, respectively. Prostaglandin D2 was widely distributed in rat brain, spinal cord and pituitary. The highest concentrations were seen in pineal gland and neurointermediate pituitary followed by anterior pituitary. Lower but significant concentrations were observed in other parts of brain, among which hypothalamus and septum showed the relatively high concentrations. 相似文献
8.
Yoshio Honma Takashi Kasukabe Motoo Hozumi Yasuko Koshihara 《Journal of cellular physiology》1980,104(3):349-357
Mouse myeloid leukemia cells (Ml) were induced to differentiate into mature macrophages and granulocytes by various inducers. The differentiated Ml cells synthesized and released prostaglandins, whereas untreated Ml cells did not. When the cells were prelabelled with [14C]arachidonate, the major prostaglandins released into the culture media were found to be prostaglandin E2, D2, and F2α in an early stage of differentiation, but the mature cells produced predominantly prostaglandin E2. The synthesis and release of prostaglandins were completely inhibited by indomethacin. Dexamethasone, a potent inducer of differentiation of Ml cells, did not induce production of prostaglandins in resistant Ml cells that could not differentiate even with a high concentration of dexamethasone. These results suggest that production of prostaglandins in Ml cells is closely associated with differentiation of the cells. Homogenates of dexamethasone-treated Ml cells converted arachidonate to prostaglandins, but this conversion was scarcely observed with homogenates of untreated Ml cells. Dexamethasone and the other inducers stimulated the release of arachidonate from phospholipids. Therefore, induction of prostaglandin synthesis during differentiation of Ml cells may result from induction of prostaglandin synthesis activity and stimulation of the release of arachidonate from cellular lipids. Lysozyme activity, which is a typical biochemical marker of macrophages, was induced in Ml cells by prostaglandin E2 or D2 alone, as well as by inducers of differentiation of the cells, but it was not induced by arachidonate or prostaglandin F2α. These results suggest that prostaglandin synthesis is important in differentiation of myeloid leukemia cells. 相似文献
9.
A method is described for the fractionation of prostaglandins and their metabolites in urine. Following acidification and extraction on Amberlite XAD-2, samples were separated by chromatography on the lipophilic anion exchanger diethyl-aminohydroxypropyl Sephadex LH-20 into fractions containing neutral compounds, monocarboxylic, dicarboxylic and polycarboxylic acids. The compounds in resulting fractions were further separated by reversed phase partition chromatography. As an application, the metabolic profiles in urine of [9β-3H]-labeled prostaglandin F2α1 and prostaglandin analogs 15-methyl-PGF2α and 16,16-dimethyl-PGF2α were investigated in the cynomolgus monkey. It was demonstrated that the resolution of individual prostaglandin metabolites by reversed phase partition chromatography was considerably simplified by initial group separation on the anion exchanger, and several metabolites were much purified. A glucuronic acid conjugate of the main metabolite of 15-methyl-PGF2α (dinor-15-methyl-PGF2α) was tentatively identified using computerized gas chromatography - mass spectrometry. 相似文献
10.
《Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism》1982,710(1):45-52
Prostaglandin synthesis by eight different structures from the rat kidney (whole cortex, cortical tubules, glomeruli, outer medulla, papilla, glomerular cultured epithelial and mesangial cells, cultured interstitial medullary cells) was measured in vitro after incubation with [14C]arachidonic acid using high-performance liquid chromatography followed by RIA with four specific anti-prostaglandin antibodies (prostaglandin E2, prostaglandin F2α, 6 keto-prostaglandin F1α, thromboxane B2). Prostaglandin production by the whole cortex and cortical tubules was very low. The order of abundance for isolated glomeruli was thromboxane B2 > prostaglandin E2 > prostaglandin F2α > 6 keto-prostaglandin F1α. Mesangial cells synthesized prostaglandin E2 at a markedly high rate, and in decreasing order: prostaglandin F2α, thromboxane B2 and 6 keto-prostaglandin F1α. The same order of abundance was observed for epithelial cells. The papilla synthesized essentially prostaglandin E2 and prostaglandin F2α, whereas the main product for the outer medulla was 6 keto-prostaglandin Fα. Cultured interstitial cells synthesized mainly prostaglandin E2 and to a lesser extent prostaglandin F2α. Unidentified peaks eluting between 6 keto-prostaglandin Fα and thromboxane B2 were also observed chiefly with glomeruli but they were absent with the medullary preparations. They disappeared after incubation with indomethacin or aspirin and represented for glomeruli the greatest percentage of conversion of [14C]arachidonic acid. These results show that the prostanoid profile varies markedly with the different regions and cells of the rat kidney. 相似文献
11.
Hideya Hayashi Seiji Ito Teruo Tanaka Manabu Negishi Hideo Kawabe Yokohama Hiromitsu Kikuko Watanabe Osamu Hayaishi 《Prostaglandins & other lipid mediators》1987,33(4)
In view of the recent finding that prostaglandin D2 is stereospecifically converted to 9α,11β-prostaglandin F2, an isomer of prostaglandin F2α, a highly specific and sensitive radioimmunoassay for 9α,11β-prostaglandin F2 was developed and applied to determine the content of this prostaglandin in various rat tissues. Antisera against 9α-11β-prostaglandin F2 were raised in rabbits immunized with the bovine serum albumin conjugate, and [3H]9α,11β-prostaglandin F2 was enzymatically prepared from [3H]prostaglandin D2. The assay detected 9α,11β-prostaglandin F2 over the range of 20 pg to 1 ng, and the antiserum showed less than 0.04% cross-section with prostaglandin F2α, prostaglandin F2β and 9β,11β-prostaglandin F2. To avoid postmortem changes, tissues were frozen in liquid nitrogen immediately after removal. The basal level of 9α,11β-prostaglandin F2 was hardly detectable in various tissues of the rat examined, including spleen, lung, liver and brain; although it was found to be 0.31 ± 0.06 ng/g wet weight in the small intestine. During convulsion induced by pentylenetetrazole, enormous amounts of prostaglandin D2 (ca. 180 ng/g wet weight) and prostaglandin F2α (ca. 70 ng/g) were produced in the brain; however, 9α,11β-prostaglandin F2 was detected neither there nor in the blood. This result demonstrates that the conversion to 9α,11β-prostaglandin F2 is a minor pathway, if one at all, of prostaglandin D2 metabolism in the rat brain. 相似文献
12.
The synthesis of deuterium- and tritium-labeled analogs of 2,3-dinor-6-keto-prostaglandin F1α and of 6,15-diketo-13,14-dihydro-2,3-dinor-prostaglandin F1α is described. These analogs were used as internal standards in the assay of the corresponding unlabeled metabolites in human urine by stable isotope dilution and combined gas chromatography-mass spectrometry. In male subjects the 24-h urinary excretion of the two metabolites was found to be 719 ± 264 and 314 ± 115 ng, respectively. The method offers a noninvasive approach to the study of prostaglandin I2 synthesis in man. 相似文献
13.
Sei-itsu Murota Mieko Kawamura 《Biochemical and biophysical research communications》1977,76(4):995-1001
Effect of various prostaglandins on the uptake of α-aminoisobutylic acid by cultured fibroblasts was studied. All the prostaglandins having an OH functional group in an intramolecular 5-membered ring showed an inhibitory effect on the amino acid uptake. The active compounds can be ranked in potency according to the values for the inhibition of the amino acid uptake per cent of control: prostaglandin F2α(53 %) >F1α(54 %) >D2(56 %) >E2(62 %) >thromboxane B2 (66 %). Thus, prostaglandin F2α was found to be the most potent inhibitor to membrane permeability and the inhibitory effect was dose dependent. The inhibition was maximal after 1 hour of exposure to prostaglandin F2α, persisted at least up to 6 hours in the presence of prostaglandin F2α. 相似文献
14.
《Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism》1981,663(2):480-490
Secondary monolayer and spinner cultures of rabbit articular chondrocytes released into the culture medium prostaglandins the synthesis of which was inhibited by sodium meclofenamate. The prostaglandins measured by radioimmunoassay were, in order of decreasing abundance, prostaglandin E2, 6-oxoprostaglandin F1α, (the stable metabolite of prostacyclin) and prostaglandin F2α. Several lines of evidence indicated that chondrocytes synthesize little if any thromboxane B2 (the stable metabolite of thromboxane A2). The presence of prostaglandins was confirmed by radiometric thin-layer chromatography of extracts of culture media incubated with [3H]arachidonic acid-labeled cells. In monolayer culture, chondrocytes synthesized immunoreactive prostaglandins in serum-free as well as serum-containing medium. Monolayer chondrocytes produced higher levels of prostaglandin E2 relative to 6-oxo-prostaglandin F1α than did spinner cells, but the latter synthesized more total prostaglandins. The identity of endogenous prostaglandins as well as those synthesized in short-term culture by rabbit cartilage slices was compared to those produced by chondrocytes in long-term culture. Chondrocytes synthesized all of the prosta-glandins found in articular cartilage. Minimal quantities of thromboxane B2 were detected in cartilage. A higher percentage of 6-oxo-prostaglandin F1α relative to other prostaglandins was found in cartilage than in either monolayer or spinner chondrocyte cultures. These results demonstrate that articular chondrocytes synthesize prostaglandins and prostacyclin. These prostaglandins may exert significant physiological effects on cartilage, since exogenous prosta-glandins depress chondrocyte sulfated-proteoglycan synthesis and may even promote proteoglycan degradation. 相似文献
15.
Shiro Ohki Katsuhiro Imaki Fumio Hirata Toshio Hanyu Nobuhiro Nakazawa 《Prostaglandins & other lipid mediators》1975,10(6):549-555
Radioimmunoassay of 5α,7α-dihydroxy-11-keto-tetranorprosta-1,16-dioic acid, main urinary metabolite of prostaglandin F2α (PGF2α), was performed using an antiserum produced in the rabbit.The antibody in 100 μ1 of 1,600-fold diluted antiserum binds with 60 picograms of metabolite.The main urinary metabolite level fell when flufenamic acid, a prostaglandin synthetase inhibitor, was given to rats. In contrast, it was significantly elevated when PGF2α was administered. 相似文献
16.
A sensitive and specific radioimmunoassay for prostaglandin D2 has been developed using its stabilized 11-methoxime derivative, which was obtained after treatment of prostaglandin D2 with methoxamine-HCl. The antiserum was obtained after injection of prostaglandin D2-methoxamine coupled to bovine serum albumin. A (125I)-Histamide prostaglandin D2-methoxamine tracer was prepared by iodination of the corresponding histamide, followed by thin layer chromatography purification. The sensitivity of the assay was 280 femtomoles per ml at 50% displacement. The cross reactivities were 15% with prostaglandin D1-methoxamine and less than 0.20% with other prostaglandins. Determination of the half-life of prostaglandin D2 in a solution containing albumin was also carried out, since it has been shown to catalyze prostaglandin D2 destruction. The unstability of this prostaglandin is due to the presence of a β-hydroxy ketone group, and all prostaglandins possessing this labile moiety could be stabilized by such a derivatization before developing a radioimmunoassay. 相似文献
17.
A series of straight phase gel chromatography systems have been developed for the separation of prostaglandin methyl esters. Using the methyl esters of prostaglandins B2, E2, F2α and F2β, the basic relationships between elution volume and the polarities of the gel, the solvent system (heptane-chloroform mixtures), and the prostaglandin have been determined. The separation of prostaglandin methyl esters with slight differences in structure has been demonstrated. Examples include oxo and hydroxy prostaglandins, hydroxy epimers, double bond isomers, prostaglandins of varying α- and ω-chain length, and 1- and 2- (5,6 cis double bond) series prostaglandins. In view of the general advantages of liquid-gel chromatography, it is suggested that these systems may be useful for isolation and purification in a number of areas in the prostaglandin field. 相似文献
18.
The behaviour of the pulmonary metabolites of prostaglandins E1, E2 and F2α were examined in several thin-layer chromatography (TLC) systems commonly used to differentiate parent prostaglandins. Although the systems chosen readily distinguished between a prostaglandin and its own metabolites, they often did not differentiate between a parent prostaglandin and the metabolites of another. In particular, 13, 14-dihydro-PGF2α was virtually indistinguishable from PGE2, and 13, 14-dihydro-PGE2 was similarly indistinguishable from PGE1, in all systems investigated. These pairs of prostaglandins could not be distinguished by bioassay on the rat stomach strip alone. Although distinction could be made by parallel assay on the rat stomach strip, chick rectum and rat colon, the differential assay obtained would not be enough to allow identification of these prostaglandins and metabolites in samples containing their mixtures. The 13, 14-dihydro-prostaglandin metabolites were also active in contracting the isolated rat uterus. The findings indicate that TLC and bioassay together may not permit identification of prostaglandins in biological fluids. 相似文献
19.
This work was undertaken to determine the ability of human prostate tissue to synthesize prostaglandin F2 alpha and to metabolize the F2 alpha to 15-keto-PGF2 alpha, 13,14-dihydro-15-keto-PGF2 alpha and 13,14-dihydro-PGF2 alpha. Surgically obtained prostate tissue was incubated with either 3H-arachidonic acid or 3H-PGF2 alpha and incorporation of tritium into PGF2 alpha or the aforementioned metabolites was determined. The identity of these substances was corroborated by gas chromatography and chemical ionization mass spectrometry. Significant increases above control levels were obtained when testosterone and lactogen were added to the incubation mixture. Maximal biosynthesis and metabolism of the prostaglandin was obtained when androgen and lactogen were present together. It is believed that androgen and lactogen produce a synergistic acceleration of PGF2 alpha synthesis and metabolism. 相似文献
20.
Metabolism of prostaglandin A. I. The kidney cortex as a major site of PGA2 degradation 总被引:2,自引:0,他引:2
The antihypertensive and natriuretic prostaglandin A2 (medullin) has been isolated and identified in rabbit renal papilla. Since PGA2, unlike prostaglandin F2 (PGE2) and prostaglandin F2α (PGF2α), is not metabolized by the lung, studies were undertaken to determine if the site of PGA2 metabolism is in the renal cortex where its primary vasodilatory and natriuretic effects occur. In experiments, homogenates of renal cortex and outer medulla were incubated with 3H-PGA2 (0.2 μc, 20 μg) at 37°C for 30 minutes. A metabolite(s) less polar than 3H-PGA2 was observed following silicic acid chromatography of acidic lipid extracts of cortical, but not outer medullary homogenates. In studies, 3H-PGA2 (2 μc, 50 μg) was injected into the renal artery of the rabbit and blood withdrawn from the ipsilateral renal vein. At least three less polar major metabolites of PGA2 were observed in the plasma within 15 seconds following injection. No appreciable 3H-PGA2 was observed in venous plasma 30 seconds following injection of 3H-PGA2. In contrast to plasma, the major urinary metabolites were more polar than PGA2. The present study reveals that PGA2 is almost completely metabolized in a single passage through the rabbit kidney suggesting this organ is a major site of PGA2 metabolism. 相似文献