RESUMO
Diagnosis of the ischemic power of epicardial stenosis with concomitant microvascular disease (MVD) is challenging during coronary interventions, especially under variable hemodynamic factors like heart rate (HR). The goal of this study is to assess the influence of variable HR and percent area stenosis (%AS) in the presence of MVD on pressure drop coefficient (CDP; ratio of transstenotic pressure drop to the distal dynamic pressure) and lesion flow coefficient (LFC; ratio of %AS to the CDP at the throat region). We hypothesize that CDP and LFC are independent of HR. %AS and MVD were created using angioplasty balloons and 90-µm microspheres, respectively. Simultaneous measurements of pressure drop (DP) and velocity were done in 11 Yorkshire pigs. Fractional flow reserve (FFR), CDP, and LFC were calculated for the groups HR < 120 and HR > 120 beats/min, %AS < 50 and %AS > 50, and additionally for DP < 14 and DP > 14 mmHg, and analyzed using regression and ANOVA analysis. Regression analysis showed independence between HR and the FFR, CDP, and LFC while it showed dependence between %AS and the FFR, CDP, and LFC. In the ANOVA analysis, for the HR < 120 beats/min and HR > 120 beats/min groups, the values of FFR (0.82 ± 0.02 and 0.82 ± 0.02), CDP (83.15 ± 26.19 and 98.62 ± 26.04), and LFC (0.16 ± 0.03 and 0.15 ± 0.03) were not significantly different (P > 0.05). However, for %AS < 50 and %AS > 50, the FFR (0.89 ± 0.02 and 0.75 ± 0.02), CDP (35.97 ± 25.79.10 and 143.80 ± 25.41), and LFC (0.09 ± 0.03 and 0.22 ± 0.03) were significantly different (P < 0.05). A similar trend was observed between the DP groups. Under MVD conditions, FFR, CDP, and LFC were not significantly influenced by changes in HR, while they can significantly distinguish %AS and DP groups.
Assuntos
Frequência Cardíaca/fisiologia , Hemodinâmica/fisiologia , Doenças Vasculares/fisiopatologia , Algoritmos , Análise de Variância , Animais , Velocidade do Fluxo Sanguíneo/fisiologia , Pressão Sanguínea/fisiologia , Capilares/fisiopatologia , Cateterismo , Circulação Coronária/fisiologia , Interpretação Estatística de Dados , Determinação de Ponto Final , Microcirculação/fisiologia , Microesferas , Análise de Regressão , SuínosRESUMO
A limitation in the use of invasive coronary diagnostic indexes is that fluctuations in hemodynamic factors such as heart rate (HR), blood pressure, and contractility may alter resting or hyperemic flow measurements and may introduce uncertainties in the interpretation of these indexes. In this study, we focused on the effect of fluctuations in HR and area stenosis (AS) on diagnostic indexes. We hypothesized that the pressure drop coefficient (CDP(e), ratio of transstenotic pressure drop and distal dynamic pressure), lesion flow coefficient (LFC, square root of ratio of limiting value CDP and CDP at site of stenosis) derived from fluid dynamics principles, and fractional flow reserve (FFR, ratio of average distal and proximal pressures) are independent of HR and can significantly differentiate between the severity of stenosis. Cardiac catheterization was performed on 11 Yorkshire pigs. Simultaneous measurements of distal coronary arterial pressure and flow were performed using a dual sensor-tipped guidewire for HR < 120 and HR > 120 beats/min, in the presence of epicardial coronary lesions of <50% AS and >50% AS. The mean values of FFR, CDP(e), and LFC were significantly different (P < 0.05) for lesions of <50% AS and >50% AS (0.88 ± 0.04, 0.76 ± 0.04; 62 ± 30, 151 ± 35, and 0.10 ± 0.02 and 0.16 ± 0.01, respectively). The mean values of FFR and CDP(e) were not significantly different (P > 0.05) for variable HR conditions of HR < 120 and HR > 120 beats/min (FFR, 0.81 ± 0.04 and 0.82 ± 0.04; and CDP(e), 95 ± 33 and 118 ± 36). The mean values of LFC do somewhat vary with HR (0.14 ± 0.01 and 0.12 ± 0.02). In conclusion, fluctuations in HR have no significant influence on the measured values of CDP(e) and FFR but have a marginal influence on the measured values of LFC. However, all three parameters can significantly differentiate between stenosis severities. These results suggest that the diagnostic parameters can be potentially used in a better assessment of coronary stenosis severity under a clinical setting.
Assuntos
Velocidade do Fluxo Sanguíneo/fisiologia , Circulação Coronária/fisiologia , Estenose Coronária/fisiopatologia , Vasos Coronários/fisiopatologia , Frequência Cardíaca/fisiologia , Análise de Variância , Animais , Pressão Sanguínea/fisiologia , Cateterismo Cardíaco , Angiografia Coronária , Modelos Animais de Doenças , Hemodinâmica , SuínosRESUMO
A three-dimensional computational model is presented for studying the efficacy of high-intensity focused ultrasound (HIFU) procedures targeted near large blood vessels. The analysis applies to procedures performed at intensities below the threshold for cavitation, boiling and highly nonlinear propagation, but high enough to increase tissue temperature a few degrees per second. The model is based upon the linearized KZK equation and the bioheat equation in tissue. In the blood vessel the momentum and energy equations are satisfied. The model is first validated in a tissue phantom, to verify the absence of bubble formation and nonlinear effects. Temperature rise and lesion-volume calculations are then shown for different beam locations and orientations relative to a large vessel. Both single and multiple ablations are considered. Results show that when the vessel is located within about a beam width (few mm) of the ultrasound beam, significant reduction in lesion volume is observed due to blood flow. However, for gaps larger than a beam width, blood flow has no major effect on the lesion formation. Under the clinically representative conditions considered, the lesion volume is reduced about 40% (relative to the no-flow case) when the beam is parallel to the blood vessel, compared to about 20% for a perpendicular orientation. Procedures involving multiple ablation sites are affected less by blood flow than single ablations. The model also suggests that optimally focused transducers can generate lesions that are significantly larger (>2 times) than the ones produced by highly focused beams.
Assuntos
Modelos Teóricos , Neoplasias/terapia , Temperatura , Terapia por Ultrassom/métodos , Ultrassom/efeitos adversos , Simulação por Computador , Humanos , Neoplasias/irrigação sanguínea , Fluxo Sanguíneo RegionalRESUMO
The peroxidase activity of the mitochondrial fraction of rat gastric mucosa was inhibited with various nonsteroidal anti-inflammatory drugs (NSAIDs) in vitro. Indomethacin was found to be more effective than phenylbutazone (PB) or acetylsalicylic acid (ASA). Mouse gastric peroxidase was also very sensitive to indomethacin inhibition. Indomethacin has no significant effect on submaxillary gland peroxidase activity of either of the species studied. Purified rat gastric peroxidase activity was inhibited 75% with 0.15 mM indomethacin showing half-maximal inhibition at 0.04 mM. The inhibition could be withdrawn by increasing the concentration of iodide but not by H2O2. NSAIDs inhibit gastric peroxidase activity more effectively at acid pH (pH 5.2) than at neutral pH. Spectral studies showed a bathochromic shift of the Soret band of the enzyme with indomethacin indicating its interaction at or near the heme part of the enzyme.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Mucosa Gástrica/enzimologia , Peroxidase/antagonistas & inibidores , Animais , Aspirina/farmacologia , Mucosa Gástrica/ultraestrutura , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Indometacina/farmacologia , Lactoperoxidase/antagonistas & inibidores , Camundongos , Camundongos Endogâmicos BALB C , Mitocôndrias/enzimologia , Fenilbutazona/farmacologia , Iodeto de Potássio/farmacologia , Ratos , Ratos EndogâmicosRESUMO
EDTA inhibits the formation of I3- from iodide catalysed by various pure peroxidases. The inhibition is concentration-dependent and chloroperoxidase (CPO) is more sensitive than horseradish peroxidase (HRP) and lactoperoxidase (LPO). EDTA is more active than EGTA or other biological chelators tested. Zn2+, Mn2+ and Co2+ are equally active in reversing the effect of EDTA on both CPO and HRP almost completely, but ineffective in the case of LPO. The effect of EDTA on HRP can be reversed by a higher concentration of iodide but not by H2O2. EDTA causes a hypsochromic change in the absorption of the Soret band of HRP at 402 nm, and iodide can reverse this effect. EDTA can effectively displace radioiodide specifically bound to HRP. It is suggested that EDTA inhibits iodide oxidation by interacting at the iodide binding site of the HRP.
Assuntos
Cloreto Peroxidase/antagonistas & inibidores , Ácido Edético/farmacologia , Peroxidase do Rábano Silvestre/antagonistas & inibidores , Iodetos/metabolismo , Lactoperoxidase/antagonistas & inibidores , Peroxidases/antagonistas & inibidores , Cinética , OxirreduçãoRESUMO
A highly active soluble peroxidase (donor: H2O2 oxidoreductase EC 1.11.1.7) has been purified from the preputial gland of the rat by hydroxylapatite chromatography, ammonium sulfate fractionation, Sephadex gel filtration and affinity chromatography on con A-Sepharose. The enzyme shows apparent homogeneity when analysed by acid and alkaline-PAGE. Its molecular, spectral, kinetic and catalytic properties were compared with those of bovine lactoperoxidase (LPO). Preputial gland peroxidase (PPO) is a glycoprotein of molecular weight of 70-73 kDa slightly lower (78 kDa) than that of LPO. Using isoelectric focussing, PPO was resolved into eight different closely spaced protein species spanning a pI range of 5.4 to 6.4, while LPO focuses into several closely spaced protein bands in the pI range 8.5-9.3. PPO is similar to LPO in its spectral (Soret) and some kinetic properties, but it differs significantly from LPO in substrate (H2O2) tolerance and substrate inactivation. PPO also differs from LPO in showing differential inactivation by SDS. Both enzymes are glycoproteins and although concanavalin A (con A) showed a variable interaction with both enzymes, wheat germ agglutinin interacted specifically with LPO only. We suggest that PPO, the secretory peroxidase of the preputial gland, differs significantly from LPO in its molecular and catalytic properties.
Assuntos
Peroxidases/isolamento & purificação , Glândulas Sebáceas/enzimologia , Sistema Urogenital/enzimologia , Animais , Eletroforese em Gel de Poliacrilamida , Guaiacol/metabolismo , Peróxido de Hidrogênio/metabolismo , Ponto Isoelétrico , Cinética , Lactoperoxidase/química , Lectinas , Peso Molecular , Peroxidases/metabolismo , Desnaturação Proteica , Ratos , Análise EspectralRESUMO
The peroxidase activity in rat gastric mucosa is inhibited after administration of glucocorticoids. The synthetic steroid dexamethasone is more potent than the naturally occurring steroids, such as cortisone or corticosterone. Almost complete inhibition of the enzyme occurs after 24 h with a single dose of 100 micrograms dexamethasone/120 g body weight. Other mitochondrial enzyme activities, like monoamine oxidase, succinic dehydrogenase and Mg2+-ATPase, remain unaltered under the same experimental condition. Submaxillary peroxidase and thyroid peroxidase activity are not inhibited by dexamethasone. Gastric peroxidase activity is increased 200-250% on the 6th day after adrenalectomy. This effect is blocked by the administration of dexamethasone. In fact, the enzyme becomes more sensitive to dexamethasone after adrenalectomy, since it is inhibited by more than 90% at the dose of 25 micrograms/120 g body weight. The inhibition by dexamethasone in normal animals is reversible. The enzyme is also inhibited after the administration of a single dose of ACTH. The apparent Km of the enzyme for H2O2 is not altered after dexamethasone treatment or after adrenalectomy. The increase in enzyme activity following adrenalectomy is not blocked by actinomycin D or by alpha-amanitin, but is prevented by puromycin or cycloheximide. After administration of dexamethasone, the iodide concentration process in the gastric mucosa is not affected, but the organification of iodide is significantly diminished.
Assuntos
Mucosa Gástrica/enzimologia , Glucocorticoides/farmacologia , Peroxidases/metabolismo , Adrenalectomia , Hormônio Adrenocorticotrópico/farmacologia , Animais , Corticosterona/farmacologia , Cortisona/farmacologia , Dexametasona/farmacologia , Hidrocortisona/farmacologia , Iodetos/metabolismo , Cinética , Masculino , RatosRESUMO
The peroxidase activity of the mitochondrial fraction of rat intestine is inhibited in vitro by non-steroidal antiinflammatory drugs (NSAIDs), such as indomethacin (IMN) and acetylsalicylic acid (ASA), the former being more potent than the latter. The peroxidase was solubilised by cetab-NH4Cl extraction and purified to apparent homogeneity by Sephadex G-150 gel filtration and affinity chromatography on Con-A Sepharose. The purified enzyme activity was 80% inhibited by 150 microM IMN and 50% by 2.67 mM ASA. IMN could also inhibit lactoperoxidase activity to the same extent but not the horseradish peroxidase activity. The inhibition of peroxidase-catalysed iodide oxidation by IMN and ASA was optimal at pH 5.5 and 4.5, respectively. Kinetic studies revealed that the inhibition by IMN was competitive with respect to iodide or guaiacol, while the inhibition by ASA was noncompetitive and reversible in nature. Studies of some structural analogues showed that indole-3-acetic acid was as effective as IMN, while salicylic acid was more potent than ASA. Spectral studies showed a small bathochromic shift of the Soret band of the enzyme by IMN, suggesting its possible interaction at or near the heme moiety. The competitive nature of IMN may be explained as due to its oxidation by the peroxidase to a product absorbing at 412 nm, the formation of which is inhibited by iodide. We suggest that IMN inhibits intestinal peroxidase activity by acting as a competitive substrate for the enzyme. As intestinal peroxidase is mainly contributed by the invading eosinophils, NSAIDs may affect the host defence mechanism by inhibiting the activity of the enzyme.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Intestinos/enzimologia , Peroxidase/antagonistas & inibidores , Animais , Bovinos , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Indometacina/farmacologia , Cinética , Oxirredução , Peroxidase/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
A protein has been solubilized and purified to homogeneity from the microsomal fraction of goat submaxillary gland. This protein can preferentially be iodinated to form triidothyronine and thyroxine with the help of submaxillary peroxidase (donor:hydrogen-peroxide oxidoreductase, EC 1.11.1.7) solubilized and purified from the same microsomal fraction. The protein can also be isolated from soluble supernatant and was found to be identical to the microsomal protein as judged by their moelcular properties as well as the formation of triiodothyronine and thyroxine. The protein has the molecular weight of 120 000 and contains two unequal subunits of molecular weight of 80 000 and 44 000. The molecular weight of the peroxidase is 72 000 and consists of a single polypeptide chain. The enzyme has the Rz value of 0.4 and is inhibited by azide and cyanide. Mersalyl, a mercurial, strongly inhibits the enzyme activity while N-ethylmaleimide cannot. The enzyme can catalyze the formation of 62 mumol of I3-/min per mg of protein at its optimun pH of 5.2. The apparent Km for H2O2 and KI is 0.16 . 10(-3) M and 1 . 10(-3) M, respectively.
Assuntos
Peroxidases/metabolismo , Proteínas e Peptídeos Salivares/metabolismo , Glândula Submandibular/análise , Tiroxina/biossíntese , Tri-Iodotironina/biossíntese , Animais , Cabras , Glândula Submandibular/enzimologiaRESUMO
Uncontrolled hydrochloric acid secretion and ulceration of the stomach mucosa due to various factors are serious global problems. Although the mechanism of acid secretion from the parietal cell is now well understood, the processes involved in gastric ulceration are still not clear. Among various causes of gastric ulceration, lesions caused by stress, alcohol consumption, Helicobacter pylori infection and due to use of nonsteroidal antiinflammatory drugs have been shown to be mediated largely through the generation of reactive oxygen species, especially the hydroxyl radical. A number of excellent drugs have proven useful in controlling hyperacidity and ulceration but their long-term use is associated with disturbing side-effects. Hence, the search is still on to find a compound possessing antisecretory, antiulcer and antioxidant properties which will serve as a therapeutic agent to reduce gastric hyperacidity and ulcers. This article describes the role of reactive oxygen species in gastric ulceration, drugs controlling them with their merits and demerits and, the role of melatonin, a pineal secretory product, in protecting against gastric lesions. In experimental studies, melatonin has been shown to be effective in reducing mucosal breakdown and ulcer formation in a wide variety of situations. Additionally, the low toxicity of melatonin supports further investigation of this molecule as a gastroprotective agent. Finally, we include a commentary on how melatonin research with respect to gastric pathophysiology can move forward with a view of eventually using this indole as a therapeutic agent to control gastric ulceration in humans.
Assuntos
Mucosa Gástrica/efeitos dos fármacos , Melatonina/farmacologia , Espécies Reativas de Oxigênio/toxicidade , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/prevenção & controle , Animais , Antiulcerosos/farmacologia , Mucosa Gástrica/fisiopatologia , Humanos , Melatonina/fisiologia , Modelos Biológicos , Úlcera Gástrica/tratamento farmacológico , Úlcera Gástrica/fisiopatologiaRESUMO
We have shown earlier that restraint-cold stress-induced gastric ulceration in rats is caused by metal ion-dependent generation of hydroxyl radical (OH.) and oxidative inactivation of the gastric peroxidase (GPO), an important H2O2 scavenging enzyme. To study the mechanism of the oxidative damage of GPO, the purified enzyme was exposed to an OH. generating system containing Cu2+, ascorbate, and H2O2. Kinetic studies indicate that the enzyme is inactivated in a time-dependent process showing saturation with respect to Cu2+ concentration. The enzyme specifically requires Cu2+ and is not inactivated by the same concentration of Fe2+, Mn2+, or Zn2+. Sensitivity to catalase indicates the critical role of H2O2 in the inactivation. Inactivation is insensitive to superoxide dismutase, suggesting no role of superoxide. The rate of inactivation is not increased in D2O excluding the involvement of singlet oxygen in the process. However, OH. scavengers such as benzoate or mannitol cannot prevent inactivation. The results indicate a plausible generation of OH. within the enzyme molecule as the cause of inactivation. Fragmentation of peptide linkage or intramolecular crosslinking, gross change of tertiary structure, or change in intrinsic tryptophan fluorescence which occurs in "global" oxidation are not evident. Inactivation is dependent on pH and from a plot of K(obs) of inactivation against pH, the controlling role of an ionizable group of the enzyme having a pka of 7.8 could be suggested, deprotonation of which favors inactivation. Amino acid analysis shows a specific loss of two lysine residues in the inactivated enzyme. Competitive kinetic studies indicate that pyridoxal phosphate, a specific modifier of the lysine residue, prevents inactivation by competing with Cu2+ for binding at the GPO. A Cu2+ binding motif consisting at least of two lysine residues exists in GPO, which specifically binds Cu2+ and generates OH.. The radical oxidizes the lysine residues and perturbs the heme environment to cause inactivation. We suggest that oxidative damage of GPO is mediated by site-specific generation of OH. and not by the OH. generated in the bulk phase.
Assuntos
Mucosa Gástrica/enzimologia , Radical Hidroxila/metabolismo , Peroxidase/antagonistas & inibidores , Úlcera Gástrica/etiologia , Estresse Fisiológico/complicações , Animais , Ácido Ascórbico/metabolismo , Ácido Ascórbico/farmacologia , Temperatura Baixa , Cobre/metabolismo , Cobre/farmacologia , Sequestradores de Radicais Livres/farmacologia , Heme/metabolismo , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Lisina/metabolismo , Oxirredução , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Restrição FísicaRESUMO
The role of the metal-catalyzed production of hydroxyl radicals (OH.) on gastric ulceration caused by restraint-cold stress in rat was studied. Stress causes a 50% increase in the thiobarbituric acid reactive species (TBARS) as a measure of the lipid peroxidation, nearly 70% increase in protein oxidation as measured by its carbonyl content and about 40% decrease in the glutathione content of the fundic stomach, suggesting oxidative damage by stress. Stress also causes a time-dependent increase in the mitochondrial superoxide dismutase activity and a decrease in the peroxidase activity, both of which correlate well with the increase in the severity of ulceration as measured by the ulcer index. Specific OH. scavengers such as benzoate or dimethylsulfoxide (DMSO) and the free radical trap such as alpha-phenyl N-tert-butyl nitrone (PBN) significantly inhibit gastric ulceration suggesting the role of OH. in this oxidative damage. Desferrioxamine (DFO), a nontoxic transition metal ion chelator, protects the mucosa against stress-ulceration dose dependently. Increased level of TBARS and the inactivation of gastric peroxidase are also prevented by DFO or by antioxidants such as glutathione or vitamin E, suggesting the critical role of metal ion and OH. in the oxidative damage. A metal-catalyzed OH. generating system constituted by Cu2+, H2O2 and ascorbate (reducing equivalent of O2-) causes inactivation of the purified gastric peroxidase in vitro, which can be effectively prevented by DFO. The stress-induced activation of the superoxide dismutase is completely blocked by pretreatment with alpha-amanitin indicating an increased synthesis of the enzyme by increased transcription of its m-RNA. Quantitative measurement indicates that stress causes a fivefold increase in the generation of OH., which correlates well with the increase in ulcer index with the progress of stress. The results indicate that the stress-induced gastric ulceration is a consequence of the oxidative damage of the gastric mucosa. This is caused by the OH. generated through the metal-catalyzed Haber-Weiss reaction between O2- and H2O2, the latter being formed by the stimulation of the superoxide dismutase and inactivation of the gastric peroxidase.
Assuntos
Mucosa Gástrica/metabolismo , Radical Hidroxila/metabolismo , Úlcera Gástrica/metabolismo , Estresse Fisiológico/metabolismo , Amanitinas/farmacologia , Animais , Quelantes/metabolismo , Quelantes/farmacologia , Temperatura Baixa , Cobre/farmacologia , Óxidos N-Cíclicos , Desferroxamina/farmacologia , Glutationa/metabolismo , Peróxido de Hidrogênio/metabolismo , Peroxidação de Lipídeos , Óxidos de Nitrogênio/farmacologia , Peroxidases/metabolismo , Proteínas/química , Proteínas/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Marcadores de Spin , Superóxido Dismutase/metabolismoRESUMO
A highly active soluble peroxidase has been identified in the preputial glands of the rat. The enzyme was detectable in the sebaceous secretion of the glands and showed catalytic properties characteristic of true peroxidase. It had a native molecular weight of around 73,000 as determined by gel-permeation studies. Immunologically the enzyme cross-reacted with an antiserum against bovine lactoperoxidase. Administration of progesterone resulted in a significant increase in the total activity of the enzyme, while testosterone and oestradiol had no such effect. The enzyme had a similar molecular weight and similar catalytic and immunological properties to rat uterine fluid peroxidase but differed markedly in respect to sensitivity to oestradiol.
Assuntos
Isoenzimas/fisiologia , Peroxidases/fisiologia , Ratos/fisiologia , Glândulas Sebáceas/enzimologia , Animais , Estradiol/metabolismo , Feminino , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Peso Molecular , Peroxidase , Peroxidases/isolamento & purificação , Peroxidases/metabolismo , Ratos Endogâmicos , Útero/metabolismoRESUMO
A method has been developed for the isolation of cells, high in iodine uptake and peroxidase activity, from the stomach and submaxillary gland of mice. The isolated cells could produce protein-bound monoiodotyrosine, di-iodotyrosine and an unknown iodocompound. The reactions were catalysed by peroxidase and were sensitive to antithyroid drugs and haemoprotein inhibitors but were insensitive to TSH. In-vitro iodination of stomach or submaxillary soluble proteins with the respective peroxidase yielded similar iodocompounds while thyroxine was produced when thyroglobulin was used instead.
Assuntos
Mucosa Gástrica/metabolismo , Isoenzimas/metabolismo , Monoiodotirosina/biossíntese , Peroxidases/metabolismo , Glândula Submandibular/metabolismo , Animais , Separação Celular , Iodetos/metabolismo , Cinética , Camundongos , Camundongos Endogâmicos , Peroxidase , Estômago/citologia , Glândula Submandibular/citologiaRESUMO
Intraperitoneal administration of mercaptomethylimidazole (methimazole), a potent antithyroid drug belonging to the thionamide group, caused a significant increase in gastric secretion both in control and pylorus-ligated mice. The drug also induced significant stimulation of gastric acid and pepsinogen secretion in both the animal systems studied. The dose-response curve indicated a nearly 10-fold increase in acid output by injection of 0.55 mg mercaptomethylimidazole per 25 g body weight. The duration profile of the drug response at the dose mentioned showed acid secretion almost at a linear rate up to 2.5 hr, after which the response decreased to some extent. Of the other antithyroid drugs of the same family, only thiourea activated acid secretion but the response was much smaller than mercaptomethylimidazole. Histamine, one of the physiological secretagogues of gastric acid secretion, was found to be less active than mercaptomethylimidazole. Mercaptomethylimidazole-induced stimulation of acid secretion could be effectively blocked by prior administration of cimetidine and completely by omeprazole and not by atropine. Verapamil and nifedipine had also some inhibitory effect. These observations indicate that mercaptomethylimidazole stimulates HCl secretion through the involvement of H2-receptor and through the functioning of the H+-K+-ATPase of the parietal cells. The bulk movement of water during increased HCl secretion was partially sensitive to cimetidine and omeprazole and was also associated with an increased secretion of Na+ and K+ in the gastric juice. This indicates that mercaptomethylimidazole also induced water transport through a separate mechanism.
Assuntos
Ácido Gástrico/metabolismo , Metimazol/farmacologia , Receptores Histamínicos H2/efeitos dos fármacos , Animais , Cimetidina/farmacologia , Masculino , Metimazol/antagonistas & inibidores , Camundongos , Camundongos Endogâmicos BALB C , Nifedipino/farmacologia , Omeprazol/farmacologia , Células Parietais Gástricas/efeitos dos fármacos , Potássio/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Piloro/efeitos dos fármacos , Receptores Histamínicos H2/fisiologia , Sódio/metabolismo , Verapamil/farmacologiaRESUMO
Mercaptomethylimidazole (MMI), a potent antithyroid drug of the thionamide group, induces both acid and pepsinogen secretion independently in control and pylorus ligated mice. The effect is dose dependent and the drug is more effective than histamine, carbachol or isoproterenol when administered by an intraperitoneal route. MMI-stimulated pepsinogen secretion could be dissociated from the acid secretion by the use of cimetidine and omeprazole which effectively block the acid secretion without affecting the pepsinogen output. Neither acid nor pepsinogen secretion by MMI is inhibited by atropine indicating a lack of muscarinic receptor involvement in both of the processes. Nifedipine and verapamil, the calcium antagonists, by inhibiting the MMI-induced acid secretion can also dissociate pepsinogen secretion from the acid secretion. Clonidine, an alpha 2-agonist, and hexobarbital, a membrane active barbiturate, also inhibit acid secretion without affecting the pepsinogen output. These data indicate that MMI induces pepsinogen secretion independent of acid secretion. Furthermore, MMI-stimulated acid secretion is not additive with that of the histamine indicating same site (H2-receptor) of action while its synergistic effect in presence of carbachol (muscarinic receptor) indicates different site of interaction of the two compounds. On the other hand, an additive effect of MMI and carbachol on pepsinogen secretion indicates that while the carbachol effect is mediated through the muscarinic receptor, MMI stimulates pepsinogen secretion through some still unknown mechanism.
Assuntos
Antitireóideos/farmacologia , Ácido Gástrico/metabolismo , Metimazol/farmacologia , Pepsinogênios/metabolismo , Piloro/efeitos dos fármacos , Animais , Carbacol/farmacologia , Histamina/farmacologia , Injeções Intraperitoneais , Isoproterenol/farmacologia , Ligadura , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pepsina A/metabolismo , Piloro/metabolismoRESUMO
Lactoperoxidase (LPO, EC 1.11.1.7; donor-H2O2 oxidoreductase) catalyses the oxidation of indomethacin, a nonsteroidal antiinflammatory drug by H2O2 as measured by time-dependent decay of indo-methacin extinction at 280 nm and concurrent appearance of stable oxidation product(s) at 412 nm. From a plot of log Vmax against varying pH of indomethacin oxidation, involvement of an ionizable group of the enzyme having pka = 5.7 could be ascertained for controlling the oxidation process. Spectral studies revealed that LPO-compound II oxidises indomethacin through one-electron transfer and is reduced to the native ferric state as shown by its spectral shift from 430 nm to 412 nm through an isosbestic point at 421 nm. The one-electron oxidation product is a nitrogen-centered free radical detected as a 5,5-dimethyl-l-pyrroline N-oxide (DMPO) adduct (alpha N = 15 G, alpha H beta = 16 G) in electron spin resonance spectroscopy. The free radical is scavenged by reaction with O2 as shown by O2 consumption sensitive to the free-radical trap, DMPO. Binding studies by optical difference spectroscopy indicate that indomethacin binds to LPO with an apparent KD value of 24.5 microM. The free energy change, delta G', for the binding is -26.3 KJ mol-1, suggesting that the interaction is favourable for oxidation. Indomethacin binding remains unaltered by a change of pH from 5.25 to 7.5, presumably because of hydrophobic interaction. The binding is competitive with resorcinol, an aromatic electron donor, showing the KD value to be as high as 100 microM. We suggest that indomethacin interacts at the aromatic donor binding site and is oxidised by one-electron transfer by LPO catalytic intermediates to stable oxidation product(s) through the formation of a free radical.
Assuntos
Anti-Inflamatórios não Esteroides/metabolismo , Indometacina/metabolismo , Lactoperoxidase/metabolismo , Animais , Anti-Inflamatórios não Esteroides/química , Sítios de Ligação , Catálise , Bovinos , Óxidos N-Cíclicos , Espectroscopia de Ressonância de Spin Eletrônica , Transporte de Elétrons , Radicais Livres/química , Radicais Livres/metabolismo , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Indometacina/química , Cinética , Oxirredução , Consumo de Oxigênio , Marcadores de Spin , TermodinâmicaRESUMO
We have reported earlier that mercaptomethylimidazole (MMI), an antithyroid drug of thionamide group, induces gastric acid secretion at least partially through the liberation of histamine, sensitive to cimetidine. Now, we show that the drug has a significant inhibitory effect on the cyclooxygenase and peroxidase activity of the prostaglandin (PG) synthetase of the gastric mucosal microsomal preparation. The effect can also be mimicked by low concentrations of H2O2. While studying the possible intracellular effect of MMI on acid secretion, a cell fraction (F3) enriched in parietal cell was isolated by controlled digestion of the mucosa with protease. This cell fraction is activated by MMI as measured by increased O2 consumption. The activation is sensitive to omeprazole, a proton-pump inhibitor, indicating that the activation is due to increased acid secretion by MMI. MMI was also found to directly inhibit the peroxidase activity of the F3 cell fraction and may thus increase the intracellular level of H2O2. The cyclooxygenase activity of the PG synthetase of the F3 cell fraction is also inhibited by MMI and the effect can be reproduced by low concentrations of H2O2. Both MMI and H2O2 can also inhibit the peroxidase activity of the PG synthetase. We suggest that in addition to the activation of the parietal cell by MMI possibly through endogenous H2O2, MMI induces acid secretion in vivo by inactivating the PG synthetase thereby inhibiting the biosynthesis of PG and removing its inhibitory influence on acid secretion so that the histamine released by MMI can stimulate acid secretion with maximum efficiency.
Assuntos
Inibidores de Ciclo-Oxigenase/farmacologia , Ácido Gástrico/metabolismo , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/enzimologia , Peróxido de Hidrogênio/farmacologia , Imidazóis/farmacologia , Prostaglandina-Endoperóxido Sintases/metabolismo , Animais , Fracionamento Celular , Ativação Enzimática/efeitos dos fármacos , Mucosa Gástrica/citologia , Mucosa Gástrica/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Microssomos/química , Microssomos/enzimologia , Células Parietais Gástricas/efeitos dos fármacos , Células Parietais Gástricas/enzimologia , Peroxidase/efeitos dos fármacos , Peroxidase/metabolismo , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacosRESUMO
Mercaptomethylimidazole (2-Mercapto-1-methylimidazole, MMI), an antithyroid drug of thionamide group, significantly activated the parietal cell for acid secretion, as evidenced by increased O2 consumption by more than 2.5-fold over the basal level. When compared, MMI-induced activation was similar to that of histamine but significantly higher than that of isobutylmethylxanthine or carbachol. Activation by MMI was not prevented by receptor blockers of the parietal cell, indicating that its effect was not mediated through the cell surface histamine-H2 receptor or the muscarinic receptor. However, the activation was almost completely blocked only by omeprazole, an established inhibitor of the terminal proton-pumping H+-K+-ATPase of the parietal cell. That MMI-induced activation was coupled with the H+ transport was further confirmed by significant increase in [14C]-aminopyrine uptake by MMI in rabbit gastric gland preparation. MMI-dependent activation of the parietal cell correlated well with the inhibition of the endogenous peroxidase activity. In vitro studies indicated that MMI irreversibly inactivated both peroxidase and catalase activity of the parietal cell in presence of H2O2. As inactivation of these H2O2-scavenging enzymes should increase accumulation of intracellular H2O2, the effect of latter was studied on acid secretion. H2O2 at a low concentration, stimulated acid secretion by sevenfold in isolated gastric mucosa, which was sensitive to omeprazole. It also significantly stimulated [14C]-aminopyrine uptake in gastric gland preparation. We suggest that MMI activated parietal cells to stimulate acid secretion by endogenous accumulation of H2O2 through inactivation of the peroxidase-catalase system.
Assuntos
Ácido Gástrico/metabolismo , Imidazóis/farmacologia , Células Parietais Gástricas/efeitos dos fármacos , Aminopirina/metabolismo , Animais , Antiulcerosos/farmacologia , Catalase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Peróxido de Hidrogênio/farmacologia , Omeprazol/farmacologia , Células Parietais Gástricas/enzimologia , Células Parietais Gástricas/metabolismo , Peroxidase/antagonistas & inibidores , Coelhos , RatosRESUMO
Dexamethasone (9 alpha-fluoro-16 alpha methyl-11 beta,17 alpha,21-trihydroxy-1,4-pregnadiene-3,20-dione-21-phosphate), a synthetic glucocorticoid, has a dual role on pepsinogen content of the gastric lumen and mucosa as measured by its peptic activity. Following stimulation the luminal peptic activity gradually decreases after 6 hr, then returns to basal levels at 18 hr and by 24 hr is inhibited by 50%. The luminal peptic activity induced by the secretory compound mercaptomethylimidazole (MMI) is also decreased. Dexamethasone effect on both basal and MMI-induced peptic activity can be reproduced by cycloheximide or puromycin, translational blockers of protein synthesis. This drug also has an independent time and dose-dependent inhibitory effect on gastric mucosal peptic activity which does not correlate with increased peptic activity of the lumen. Dexamethasone appears to be more effective than hydrocortisone and corticosterone in inhibiting the basal peptic activity of both lumen and mucosa. The inhibitory effect of this drug on tissue peptic activity is not mediated through induction of any inhibitory protein as evidenced by the insensitivity of the effect to actinomycin D. Studies on [14C]phenylalanine incorporation into gastric protein indicate that the effect of dexamethasone on tissue pepsinogen content is not due to a generalized block of protein synthesis.