RESUMO
The drug trans-1,4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944) almost completely inhibited the conversion of [2-14C] mevalonic acid, dihydro[14C]lanosterol, 4,4-dimethyl-5alpha-[2-3H2]cholesta-8,14-dien-3beta-ol and 4,4-dimethyl-5alpha-[2-3H2]cholest-8(14)-en-3beta-ol to 5alpha-cholest-7-en-3beta-ol and cholesterol by cell-free systems of rat liver. With the first three precursors, the inhibition was accompanied by an accumulation of radioactive 5alpha-cholesta-8,14-dien-3beta-ol, but this material could not be detected during inhibition of cholesterol biosynthesis from 4,4-dimethyl-5alpha-[2-3H2] cholest-8(14)-en-3beta-ol. Regardless of the nature of the precursor, trans-1,4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride did not result in the accumulation of any delta5,7 sterols. Non-radioactive 5alpha-cholest-8(14)-en-3beta-ol inhibited the conversion of dihydro[14C]lanosterol to 4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol. Carbon monoxide resulted in a decrease in the rate of conversion of dihydro[14C]lanosterol to 4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol but had no effect on the rate of conversion of 4,4-dimethyl-5alpha-[2-3H2]cholesta-8,14-dien-3beta-ol to 5alpha-cholest-7-en-3beta-ol and cholesterol suggesting that cytochrome P-450 is involved neither in the oxidative removal of the 4-methyl groups nor in the oxidative introduction of the delta5 bond during cholesterol biosynthesis. In addition, the process of cholesterol and 5alpha-cholest-7-en-3beta-ol biosynthesis from 4,4-dimethyl-5alpha-[2-3H2]cholest-8(14)-en-3beta-ol was inhibited by carbon monoxide at a stage after the formation of 5alpha-cholest-8(14)-en-3beta-ol.
Assuntos
Monóxido de Carbono/farmacologia , Colesterol/biossíntese , Cicloexanos/análogos & derivados , Fígado/metabolismo , Esteróis/metabolismo , Animais , Compostos de Benzil/farmacologia , Cicloexanos/farmacologia , Lanosterol/análogos & derivados , Lanosterol/metabolismo , Fígado/efeitos dos fármacos , Metilaminas/farmacologia , Ácido Mevalônico/metabolismo , Ratos , Relação Estrutura-AtividadeRESUMO
Incubation of rat liver microsomal fraction in the presence of increasing concentration of a serum preparation and the re-isolation of the treated microsomal vesicles resulted in a progressive increase in the concentration of non-esterified cholesterol, a progressive decrease in the activity of hydroxymethylglutaryl-CoA reductase and progressive changes in the characteristics of the Arrhenius plots of the enzyme. The changes in the characteristics of the Arrhenius plots of the enzyme in the serum-treated preparations are consistent with a progressive increase in the concentration of non-esterified cholesterol in the environment of the hydroxymethylglutaryl-CoA reductase in endoplasmic reiticulum vesicles. The serum-treated preparations with high non-esterified cholesterol content showed a constant activation energy between 37 and 20 degrees C, whereas the enzyme in the non-treated microsomal fraction, the buffer-treated and the lipoprotein-deficient serum-treated preparations showed breaks in the activation energy at about 29 degrees C. The microsomal fraction from rats fed on the standard, cholesterol- or cholestyramine-supplemented diet showed considerable differences in the activity of hydroxymethylglutaryl-CoA reductase and differences in the characteristics of their Arrhenius plots. However, the incubation of the microsomal fraction from the rats in the three experimental conditions with ATP and Mg2+ and the further incubation of the inactivated enzyme with a preparation of cytosolic phosphoprotein phosphatase resulted in Arrhenius plots with similar characteristics to those of the corresponding original microsomal fraction. These results suggest that changes in the concentration of non-esterified cholesterol in the endoplasmic reticular membrane are responsible for the differences in the activity of hydroxymethylglutaryl-CoA reductase in the microsomal fraction from the rats in these dietary conditions.
Assuntos
Colesterol/metabolismo , Hidroximetilglutaril-CoA Redutases/metabolismo , Microssomos Hepáticos/enzimologia , Animais , Colesterol na Dieta , Resina de Colestiramina/farmacologia , Cinética , Lipoproteínas/fisiologia , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Ratos , Ratos Endogâmicos , TermodinâmicaRESUMO
The rates of incorporation of (14C) cholesterol into cholesteryl esters and 5-cholestene-3beta,7alpha-diol (7alpha-hydroxycholesterol) by rat liver microsomes, measured under conditions in which esterification and 7alpha-hydroxylation are varied independently, indicated that cholesterol is the substrate for cholesterol 7alpha-hydroxylase. The specific activities of cholesteryl esters and 7alpha-hydroxycholeste ol in incubations of microsomes labelled with (14C)cholesterol in vitro or in vivo suggest that 7alpha-hydroxycholesterol and esterified cholesterol are not derived from the same pool of free cholesterol.
Assuntos
Colesterol 7-alfa-Hidroxilase/metabolismo , Microssomos Hepáticos/enzimologia , Esteroide Hidroxilases/metabolismo , Trifosfato de Adenosina/farmacologia , Animais , Coenzima A/farmacologia , Ésteres , Cinética , NADP/farmacologia , Oxirredução , Ratos , Relação Estrutura-Atividade , Fatores de TempoRESUMO
The effect of high density lipoproteins (HDL) on the stability and clearance of injected liposomes was investigated under conditions of abnormal lipoprotein metabolism in vivo. Small unilamellar liposomes composed of phosphatidylcholine and containing quenched carboxyfluorescein were injected intravenously or intraperitoneally into normal mice or mice previously made lipoprotein deficient with 4-aminopyrazolo[3,4d]pyrimidine (4-APP). As evidenced from quenched carboxyfluorescein values in the blood, levels of stable liposomes in the circulation were increased and clearance rates reduced considerably in lipoprotein-deficient animals indicating increased bilayer integrity. This was confirmed by the demonstration that transfer of liposomal phosphatidylcholine to HDL, occurring in the presence of normal mouse plasma, was virtually abolished in the presence of plasma from lipoprotein deficient mice. The role of other lipoprotein species in destabilizing liposomes was also investigated. Plasma from lipoprotein-deficient mice was supplemented with increasing amounts of HDL, LDL + IDL or VLDL (to cover the physiological range of lipoprotein concentrations in mouse blood) prior to the addition of phosphatidylcholine liposomes, and incubated at 37 degrees C. It was shown that among the lipoprotein species studied only HDL was detrimental to liposomal stability under the conditions employed. Our results indicate that use of liposomal drugs in the treatment of patients must take into account HDL fluctuations in their blood as these could after liposomal membrane permeability to the drugs and thus upset therapeutic efficiency.
Assuntos
Lipoproteínas/deficiência , Lipossomos/administração & dosagem , Fosfatidilcolinas/metabolismo , Animais , Estabilidade de Medicamentos , Fluoresceínas , Humanos , Cinética , Lipoproteínas/sangue , Masculino , CamundongosRESUMO
The activity and the kinetic properties of hydroxymethylglutaryl-CoA reductase and of acyl-CoA:cholesterol acyltransferase in the liver microsomal fraction have been compared between rats fed on either unsaturated or on saturated fat. When rats wre fed for 12h on a compounded diet containing either safflower seed oil or tristearin the composition of the fatty acyl chains of the microsomal phospholipids was shown to be relatively more unsaturated in the rats that received the unsaturated fat. The activity of hydroxymethylglutaryl-CoA reductase in the microsomal fraction was considerably reduced in rats fed on compounded diet containing unsaturated fat whereas this dietary condition resulted in a considerable increase in the activity of acyl-CoA:cholesterol acyltransferase. Similar effects were observed after feeding rats for 12 h on a commercial diet supplemented with either safflower seed oil or with tristearin. The addition of 2% cholesterol to the fat-supplemented diets resulted in both cases in a decrease in hydroxymethylglutaryl-CoA reductase and an increse in acyl-CoA:cholesterol acyltransferase activity as compared with the corresponding values from the rats fed on the fat-supplemented diets with no cholesterol. The Arrhenius plots of hydroxymethylgutaryl-CoA reductase in the microsomal fraction from rats fed on fat-supplemented commercial diet for 12 h showed breaks in the activation energy at 29.6 degrees C for the preparations from rats fed on tristearin and 28 degrees C for those from rats fed on safflower seed oil. The activation energy of the enzyme was lower above and higher below the break for the preparations from rat fed on the unsaturated fat-supplemented diet. Similar differences were obtained from the comparison of the Arrhenius plots in the preparations from rats fed on saturated fat and those in the preparations from rats fed on unsaturated fat when the diet was compounded and given to the animals for 36 h. The addition of 2% cholesterol to the commercial diet supplemented with either saturated or unsaturated fat resulted in Arrhenius plots with a constant activation energy between 37 and 22 degrees C for the enzyme in microsomal preparations from both groups of rats. The apparent Km value for hydroxymethylglutaryl-CoA was lower for the reductase in microsomal preparations from rats fed on the unsaturated fat as compared with that for the enzyme in microsomal preparations from rats fed on saturated fat. There was also a decrease in the apparent Km value for oleic acid for the acyltransferase from rats fed on unsaturated fat as compared with that for the enzyme in the microsomal preparation from the rats fed on saturated fat. The results of the present study are consistent with higher concentration of free cholesterol in endoplasmic reticular membrane in the environment of the reductase and that of acyltransferase following the administration of dietary unsaturated fat as compared with that following the administation of saturated fat.
Assuntos
Aciltransferases/metabolismo , Gorduras na Dieta/farmacologia , Hidroximetilglutaril-CoA Redutases/metabolismo , Microssomos Hepáticos/enzimologia , Esterol O-Aciltransferase/metabolismo , Animais , Colesterol/metabolismo , Retículo Endoplasmático/metabolismo , RatosRESUMO
The administration of mevalonic acid to rats by intravenous injection resulted in a dose- and time-dependent increase in the activity of cholesterol 7alpha-hydroxylase in the liver microsomal fraction, a decrease in the microsomal activity of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and no significant change in the activity of acyl-coenzyme A:cholesterol acyltransferase or in the concentration of free and of esterified cholesterol in the liver microsomal fraction. However, the increased hepatic cholesterogenesis that follows the injection of mevalonic acid resulted in an increase of the size of the intracellular pool of cholesterol that is in the environment of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and acts as substrate for cholesterol 7alpha-hydroxylase. The administration of mevalonic acid to rats by stomach tube resulted in an increase in the activity of cholesterol 7alpha-hydroxylase and of acyl-coenzyme A:cholesterol acyltransferase and in the concentration of cholesterol esters in the liver microsomal fraction, while there was a considerable decrease in the activity of 3-hydroxy-3-methylglutaryl-coenzyme A reductase.
Assuntos
Aciltransferases/metabolismo , Colesterol 7-alfa-Hidroxilase/metabolismo , Colesterol/metabolismo , Hidroximetilglutaril-CoA Redutases/metabolismo , Ácido Mevalônico/farmacologia , Microssomos Hepáticos/metabolismo , Esteroide Hidroxilases/metabolismo , Esterol O-Aciltransferase/metabolismo , Animais , Colesterol/biossíntese , Ésteres do Colesterol/metabolismo , Masculino , Ácido Mevalônico/administração & dosagem , Ratos , Fatores de TempoRESUMO
1. The effect of a portacaval anastomosis on the activities of hepatic enzymes related to cholesterol metabolism was investigated in rats. 2. Portacaval anastomosis led to a fall in body weight and liver weight/body weight ratio, and to a rise in the activities of hydroxymethylglutaryl-CoA reductase and cholesterol 7alpha-hydroxylase per g of liver. The net effect was to maintain a normal activity of both enzymes per 100 g of rat. Diurnal rhythm in the activities of both enzymes was maintained after portacaval anastomosis. 3. The rate of excretion of total bile acids, per 100 g of rat, in bile fistula rats was not significantly decreased by portacaval anastomosis.
Assuntos
Ácidos e Sais Biliares/metabolismo , Colesterol/metabolismo , Fígado/metabolismo , Derivação Portocava Cirúrgica , Animais , Fístula Biliar , Colesterol 7-alfa-Hidroxilase/metabolismo , Ritmo Circadiano , Sistema Enzimático do Citocromo P-450/metabolismo , Hidroximetilglutaril-CoA Redutases/metabolismo , Masculino , Oxirredutases/metabolismo , Ratos , Tirosina Transaminase/metabolismoRESUMO
1. The effect of portacaval anastomosis on the metabolism of plasma lipoproteins was investigated in rats. 2. When compared with sham-operated pair-fed controls, plasma high density lipoprotein cholesterol concentration was decreased, plasma low density lipoprotein cholesterol concentration was increased and plasma total cholesterol concentration was unchanged in the portacaval anastomosis rats. Maximal incorporation of [14C]leucine into the total circulating mass of protein was decreased in the very low density lipoprotein and high density lipoprotein fractions and, possibly, in the low density lipoprotein fraction, but there was no change in maximal incorporation into albumin. It is concluded that portacaval anastomosis diminishes the rate of synthesis of high density lipoprotein and very low density lipoprotein proteins and, possibly, of low density lipoprotein proteins.
Assuntos
Lipoproteínas/sangue , Derivação Portocava Cirúrgica , Animais , Peso Corporal , Colesterol/sangue , Leucina/metabolismo , Fígado/metabolismo , Masculino , Tamanho do Órgão , Ratos , Albumina Sérica/metabolismoRESUMO
The incubation at 37 degrees C of rat-liver microsomal fraction followed by re-isolation of the treated microsomal vesicles results in a time-dependent increase in the activity of acyl-CoA:cholesterol acyltransferase. The rate of this increase was higher in the microsomal fraction from rats fed cholesterol-supplemented diet or starved overnight as compared with that in the microsomal fraction from rats fed standard diet. The presence of a plasma membrane preparation in the incubation mixture also resulted in a time-dependent increase in acyl-CoA:cholesterol acyltransferase activity at a rate that was dependent on the concentration of plasma membranes. During the incubation of the microsomal fraction in the presence of phosphatidylcholine liposomes, cholesterol is transferred from the microsomal to liposomal vesicles. This transfer followed first-order kinetics with respect to cholesterol concentration in the donor with a rate that increased with the concentration of liposomes in the incubation mixture. The presence of phospholipid was also associated with a decrease in the activity of the acyltransferase that was related to the concentration of phospholipid in the incubation mixture. The incubation of the microsomal fraction in the presence of phosphatidylcholine-cholesterol liposomes resulted in a time-dependent and concentration-dependent transfer of liposomal cholesterol to the microsomal fraction and the acyltransferase substrate pool. The measurement of the rate of transfer of liposomal cholesterol to the microsomal vesicles and to the acyltransferase substrate pool at various temperatures showed that activation energies for the two processes are similar. Similar to these various was also the activation energy for the increase in acyl-CoA:cholesterol acyltransferase activity due to preincubation in the absence of artificial membrane vesicles. The present results suggest that there is, under the present conditions, a time-dependent and temperature-dependent flow of cholesterol from plasma membranes to the acyltransferase substrate pool and that this flow is either diverted in the presence of phospholipid liposomes or increased in the presence of cholesterol-phospholipid liposomes.
Assuntos
Aciltransferases/metabolismo , Colesterol/metabolismo , Fígado/metabolismo , Microssomos Hepáticos/metabolismo , Esterol O-Aciltransferase/metabolismo , Animais , Transporte Biológico , Membrana Celular/metabolismo , Cinética , Bicamadas Lipídicas , Masculino , Fosfatidilcolinas , Ratos , Ratos Endogâmicos , TrítioRESUMO
The subcellular site of accumulation of non-esterified cholesterol and sphingomyelin in lipid-laden livers and spleens from rats given multiple intravenous injections with liposomes made up of these lipids were investigated by morphological and biochemical techniques. The subcellular fractionation of liver homogenates from cholesterol-sphingomyelin treated rats followed by lipid and enzymatic analyses of the fractions revealed that most of the accumulating lipid was present in very low density lysosomes floating in the post-microsomal supernatant fraction. The low density lysosomes exhibited good latency and had a very much lower relative activity of sphingomyelinase compared with values for N-acetyl-beta-glucosaminidase. Multiple injections of sphingomyelin-cholesterol liposomes resulted in splenomegaly. The spleen homogenates of the treated rats showed a many-fold increase in the concentration of sphingomyelin, of non-esterified cholesterol and in the activity of N-acetyl-beta-glucosaminidase over corresponding values for rats injected with saline. Electron microscopy of liver and spleen sections from treated rats revealed distinctive polymorphic intracellular inclusions bound by a membrane and containing numerous osmiophilic bodies. Structures identical to the storage inclusions seen in fixed liver sections from treated rats were also seen by electron microscopy in the post-microsomal fraction of these livers. The results suggest that sphingomyelin and non-esterified cholesterol accumulate in lysosomes when they occur in cells in excess of that structurally associated with cellular membranes.
Assuntos
Acetilglucosaminidase/metabolismo , Colesterol/metabolismo , Hexosaminidases/metabolismo , Fígado/metabolismo , Lisossomos/metabolismo , Inibidores de Fosfodiesterase , Esfingomielina Fosfodiesterase/antagonistas & inibidores , Esfingomielinas/metabolismo , Animais , Fracionamento Celular , Fígado/ultraestrutura , Lisossomos/ultraestrutura , Masculino , Microscopia Eletrônica , Diester Fosfórico Hidrolases , Ratos , Ratos Endogâmicos , Frações Subcelulares/metabolismoRESUMO
Factor VII coagulant activity (VIIc) is considerably higher in rabbits fed a 1% cholesterol-supplemented diet than in rabbits fed a standard diet. This increase was statistically significant 4-6 days from the beginning of treatment and rose to about 300% during the 100 days of treatment. Treatment is also associated with a 20-fold increase in plasma cholesterol concentration with the major fraction of excess cholesterol associated with the very low and intermediate density lipoprotein fractions. In both groups of rabbits, the direction and extent of variation in VIIc generally coincided with variation in cholesterol, so that over time there were significant and positive correlations between plasma cholesterol concentration in both the rabbits fed the standard diet and the rabbits fed the cholesterol-supplemented diet. The increase in VIIc was due to a higher proportion of the more active alpha VIIa in the plasma of hypercholesterolaemic rabbits rather than to an increase in the concentration of the single-chain protein. The plasma concentration of factor X and prothrombin had increased in the hypercholesterolaemic rabbits by 10 days from the beginning of treatment and both proteins were maintained at 150-200% of the concentrations in the plasma of rabbits fed the standard diet. However, these differences were only seen when the factor X and prothrombin were assayed using synthetic substrates. The specific coagulation assays for these two factors revealed no differences between the groups of animals up to 100 days.
Assuntos
Antígenos/metabolismo , Fator VII/imunologia , Hipercolesterolemia/sangue , Animais , Coagulação Sanguínea , Colesterol/sangue , Colesterol na Dieta/administração & dosagem , Fator VII/metabolismo , Fator VIIa , Fator X/metabolismo , Hipercolesterolemia/etiologia , Masculino , Protrombina/metabolismo , CoelhosRESUMO
Cholesterol and triglyceride in the various lipoprotein fractions were determined in five patients without functional lipoprotein lipase (LPL) while on their habitual therapeutic diet of 'low fat' content (20-25 g/day). They were also studied following 3 days on either a 'minimal fat' diet (< 15 g/day) or a 'moderate fat' diet (45-50 g/day). Values obtained were compared with the respective levels measured in five control subjects on a 'normal fat' (70-90 g/day) diet. The patients had hypertriglyceridaemia (type V hyperlipoproteinaemia) under all dietary conditions. Cholesterol and triglyceride levels in plasma and in the chylomicron fraction increased in the patients with increasing dietary fat. In the very low density lipoprotein (VLDL) fraction from the patients, triglyceride levels also increased with the dietary fat intake, but cholesterol levels were similar under all dietary conditions. In the patients, cholesterol concentrations in the low (LDL) and high density (HDL) lipoprotein fractions were significantly lower than the respective levels in controls, but the ratio of cholesterol to triglyceride levels in both of these lipoprotein fractions decreased with the dietary fat intake. VLDL apolipoprotein B-100 (apo B-100) pool size was similar in the patients on the two test diets (P = 0.95) and 3.5-fold higher than in five healthy volunteers on a normal fat diet. Using a stable isotope enrichment method, the kinetics of apo B-100 were investigated in the patients under the last two dietary conditions. The fractional and absolute secretion rates of the apolipoprotein in the patients did not vary with fat intake, but fractional secretion rates were significantly lower and the absolute secretion rates were significantly higher in the patients than the respective values in the controls. These results are consistent with the hypothesis that in the absence of LPL activity the metabolism of chylomicron and VLDL particles in the circulation results in triglyceride-rich LDL and HDL particles that are taken up by the liver at increased rates, thus reducing the plasma LDL and HDL cholesterol concentrations, whereas the products of hydrolysis of these particles induce an increased rate of synthesis of triglyceride and an increased rate of secretion of VLDL apo B-100.
Assuntos
Apolipoproteínas B/análise , Lipase Lipoproteica/deficiência , Lipoproteínas/sangue , Triglicerídeos/sangue , Adulto , Apolipoproteína B-100 , Colesterol/sangue , Feminino , Humanos , Hiperlipoproteinemia Tipo V/sangue , Hiperlipoproteinemia Tipo V/enzimologia , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/sangue , Masculino , Pessoa de Meia-IdadeRESUMO
A community survey of factor VII coagulant activity (VIIc) and the lipoprotein profile in non-fasting plasma of middle-aged men in NW London was undertaken to search for the determinants of VIIc in the general community. The data demonstrates that associations between VIIc and the plasma concentrations of cholesterol and of triglycerides previously shown in the general population can be explained by the strong and positive associations between VIIc and the large lipoprotein particles, chylomicrons, VLDL and IDL. Consistent with the possibility that the concentration of large lipoproteins determines the in vivo reactivity of factor VII, the association between VIIc and the ratio of lipid in the d greater than 1.019 fraction to the total plasma lipid was also highly significant but negative. The observed correlations between VIIc and lipoproteins smaller than VLDL may be the product of the interrelations that exist between the lipoprotein fractions in plasma. However, the associations between VIIc and the chylomicron lipid concentrations are especially strong when allowance is made for the considerable bias towards zero in the observed correlation, due to large within-person variance in chylomicron concentration.
Assuntos
Fator VII/metabolismo , Hiperlipoproteinemias/sangue , Lipoproteínas/sangue , Fatores Etários , Coagulação Sanguínea , Quilomícrons/sangue , Fator VII/fisiologia , Humanos , Lipoproteínas VLDL/sangue , Masculino , Pessoa de Meia-IdadeRESUMO
A high factor VII coagulant activity (VIIc), a marker of increased risk of coronary heart disease, is frequently found in types IIb and IV hyperlipidaemia, but its cause is not fully understood. Factor VII can be activated by factor XIIa, generated from factor XII upon activation of the contact system of coagulation. Ten patients with familial lipoprotein-lipase (LPL) deficiency and 10 healthy control subjects were therefore compared to explore the hypothesis that high concentrations of unesterified fatty acids (UFA), released from triglyceride-rich lipoproteins by LPL, are a source of factor XII activation and hence the increased VIIc that is observed post-prandially and in non-LPL-deficient hypertriglyceridaemic states. Mean plasma cholesterol and triglyceride concentrations were, respectively, 1.5- and 19-fold higher in the patients than controls, due to increases in very-low-density lipoproteins and chylomicrons. The concentration and composition of plasma UFA were similar in both groups. In conformity with the hypothesis, VIIc was not increased in the LPL-deficient group, despite their massive hypertriglyceridaemia. Furthermore, when the patients' plasma was treated with LPL, factor XII was activated promptly and substantially, whereas no similar effect was observed in the controls. These results suggest that high concentrations of circulating triglyceride-rich lipoproteins will increase VIIc in the presence of LPL.
Assuntos
Fator XII/fisiologia , Lipólise , Lipase Lipoproteica/deficiência , Lipoproteínas/metabolismo , Triglicerídeos/metabolismo , Adulto , Fator VII/metabolismo , Fator XII/metabolismo , Ácidos Graxos não Esterificados/sangue , Ácidos Graxos não Esterificados/classificação , Feminino , Humanos , Lipoproteínas/sangue , Masculino , Pessoa de Meia-IdadeRESUMO
In order to evaluate whether Hageman factor (XII) is increased in survivors of myocardial infarction and whether this in turn influences factor VII coagulant activity (VIIc), we examined the coagulation and lipoprotein profiles in 82 subjects, 51 of whom had a definite history of myocardial infarction and 31 healthy volunteers invited from a local general practice register for a cardiovascular screen. Both serum cholesterol (P = 0.03) and plasma fibrinogen levels (P = 0.02) were significantly elevated in cases compared with controls. There were no significant differences in coagulant activities, and in particular factor XII concentration was not significantly different between groups. Furthermore, in 47 of the subjects, 28 of whom had a history of myocardial infarction, a more detailed analysis, including measurement of VIIc after overnight incubation of plasma at 4 degrees C, was undertaken. Approximately half the subjects in either group showed some evidence of activation, though history of myocardial infarction was not in itself a significant predictor of this. All measures of XII concentration related positively to VIIc after cold activation, the strongest being the measure of amidolytic activity following activation of factor XII (XIIAm) (r = 0.5, P < 0.01). In addition, XIIa, a measure of activity due to enzymes derived from factor XII, related strongly to many of the measured lipoprotein variables, particularly VLDL cholesterol and triglycerides, supporting the hypothesis that negatively charged molecules such as free fatty acids on larger lipoprotein particles provide the contact surface necessary to activate factor XII. The findings confirm the importance of this alternative pathway in leading to activation of factor VII.
Assuntos
Fator XII/análise , Infarto do Miocárdio/sangue , Idoso , Antígenos/análise , Colesterol/sangue , Fator VII/análise , Fibrinogênio/análise , Humanos , Lipoproteínas/sangue , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
Nine adults took two 7-day diets of standardised energy and total fat content, but with a dietary polyunsaturated/saturated fat ratio of less than 0.3 and greater than 3.0 respectively, while adhering to their daily routine. Blood was drawn on 6 occasions between 09.00 and 22.45 h on the final day of each dietary period for factor VII activity (VIIc), factor VII antigen (VIIag) and lipoprotein lipid concentrations. Diurnal variation was described for each variable in terms of its deviation from the individual's daily mean value at each time point across the day. Plasma triglyceride remained low until after the midday meal, whereafter a marked rise was sustained into the later evening. Plasma VIIc declined until early afternoon, but showed a marked rise in the late afternoon. Plasma VIIag showed no significant diurnal variation. Changes in plasma triglyceride concentration during the day were related positively to changes in VIIc about 160 min later, but not to VIIc at other time points. This effect of postprandial triglyceridaemia on VIIc persisted after allowance for the effect of VIIag on VIIc. Dietary fat composition did not influence VIIc or VIIag. The results suggested an acute but evanescent effect of triglyceride-rich lipoproteins on the reactivity of factor VII, irrespective of their lipid core composition.
Assuntos
Gorduras na Dieta/administração & dosagem , Fator VIIa/metabolismo , Triglicerídeos/sangue , Antígenos/análise , Ritmo Circadiano , Fator VII/análise , Fator VII/imunologia , Feminino , Humanos , Lipoproteínas/sangue , MasculinoRESUMO
Diet was measured by 5-day weighed inventory to search for an association between fat intake in the general population and factor VII coagulant activity (VIIc), a strong predictor of coronary heart disease. Of 275 men aged 40-59 years registered with a medical practice, 203 (74%) participated and 170 (62%) provided a satisfactory record. After allowance for the increase in fat intake with body size, a statistically significant and positive association was found between dietary fat and VIIc (r = 0.18; P less than 0.05). The correlation coefficient was increased to 0.24 when adjusted for the effect of day-to-day variability in individual fat intake, thereby providing an improved estimate of the true strength of association. The mean difference in VIIc of 12% of standard between men in the highest and lowest quarters of the distribution of fat intake was similar to that reported between men experiencing coronary heart disease and those remaining free. The results support previous experimental fat-feeding studies and suggest that a high fat diet has adverse consequences for blood coagulability and coronary thrombosis.
Assuntos
Antígenos/análise , Doença das Coronárias/etiologia , Gorduras na Dieta/administração & dosagem , Fator VII/imunologia , Adulto , Consumo de Bebidas Alcoólicas , LDL-Colesterol/sangue , Carboidratos da Dieta/administração & dosagem , Gorduras na Dieta/sangue , Gorduras na Dieta/metabolismo , Proteínas Alimentares/administração & dosagem , Fator VII/análise , Fibrinogênio/análise , Humanos , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
The contribution of the various components of the contact system in the generation of factor XIIa (FXIIa) and of kallikrein (KRN) on an electronegative surface and the release of the generated enzymes to the bulk phase was examined in mixtures of normal human plasma and plasmas congenitally deficient in these components. The incubation of normal human plasma in the presence of sulphatide vesicles (40 microM) resulted in a fast generation of amidolytic activities due to FXIIa and to KRN followed by slower first-order inactivation rates of FXIIa (k'FXIIa) and of KRN (k'KRN) due to the presence of esterase inhibitors. Variation of the levels of factor XII (FXII), over a wide range, showed little effect on levels of FXIIa and of KRN but no activities were detected in 100% FXII-deficient plasma. The variation of prekallikrein (PKRN) concentration showed little effect on the generation of FXIIa but the generation of KRN declined linearly with the decrease in the level of PKRN. No activities were detected on treatment of PKRN-deficient plasma. The variation in the concentration of high molecular weight kininogen (HK) showed effects on FXIIa and KRN that were qualitatively similar to those seen on variation of PKRN but 100% HK-deficient plasma generated considerable activities of both FXIIa and KRN. The variation in the concentration of factor XI (FXI) showed no effect on the generation of FXIIa, whereas KRN levels increased linearly with the contribution of FXI-deficient in normal plasma. The present results suggest that the contiguous binding of FXIIa, FXII, PKRN-HK and FXI-HK onto the electronegative surface induces a rapid generation of FXIIa and KRN. The bound PKRN-HK complex prevents the release of generated FXIIa and therefore further binding and activation of FXII from the bulk phase. Consequently, the turnover of FXII is independent of its levels in the bulk phase and is rather related to the concentration of contact surface. The generated KRN is also protected by HK. However, since the enzyme responsible for the activation of PKRN-HK is FXIIa, the levels of generated KRN are positively related to the concentration of substrate.
Assuntos
Fator XIIa/biossíntese , Animais , Coagulação Sanguínea/fisiologia , Bovinos , Eletroquímica , Fator XI/metabolismo , Fator XII/metabolismo , Humanos , Técnicas In Vitro , Cinética , Cininogênio de Alto Peso Molecular/sangue , Pré-Calicreína/metabolismo , Sulfoglicoesfingolipídeos/metabolismo , Propriedades de SuperfícieRESUMO
The incubation of purified human factor XII (Hageman factor [HF]) in the presence of long-chain saturated fatty acids (FA) like stearate (C-18) or behenate (C-22) resulted in a time-dependent increase of amidolytic activity. The HF autoactivation progress curves were sigmoidal. The first order rate for the initial period was constant; this was followed by a period of decreasing rate and a plateau of zero rate. These progress curves were similar to those obtained on the incubation of HF in the presence of sulphatide vesicles or dextran sulphate. The initial rate of autoactivation of HF was dependent on the FA concentration of contact surface and increased with increasing concentration of HF. At constant concentration of contact surface and varying concentration of HF, autoactivation rates in the presence of behenate, sulphatide vesicles or dextran sulphate followed Michaelis-Menten kinetics. The Km values for all three contact surfaces were above the physiological plasma concentration of HF whereas the catalytic efficiency in the presence of behenate (0.034 microM-1s-1) was about 2/3 of that in the presence of sulphatide vesicles (0.053 microM-1s-1) and considerably higher than that in the presence of dextran sulphate (0.004 microM-1s-1). Long-chain saturated FA bound to human serum albumin at the high- or low-affinity sites are ineffective, whereas the crystalline non-bound stearate or behenate provided a potent contact surface.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Sulfato de Dextrana/farmacologia , Fator XII/metabolismo , Ácidos Graxos/farmacologia , Sulfoglicoesfingolipídeos/farmacologia , Ácidos Graxos/química , Humanos , Concentração de Íons de Hidrogênio , Cinética , Lipossomos , Albumina Sérica/químicaRESUMO
The amidolytic activity of enzymes derived from factor XII (XIIa) was 3-fold higher in plasmas collected during pregnancy than from control subjects. Factor VII coagulant activity (VIIc) and XIIa increased in both kinds of plasmas on incubation on ice for 24 h (cold activation). These increases could be attributed to the decreased potency of C1 inhibitor (C1INH). However, variations in the concentration of C1INH and of factor XII could not explain the differences in VIIc and in XIIa between late pregnancy and control plasmas following cold activation under the same conditions. It is concluded that in vitro the increased amount of contact surface in the late pregnancy plasma promotes a higher rate of generation of XIIa and consequently a higher rate of activation of factor VII. The increased amount of contact surface could also be responsible for the increased concentration of XIIa in non-treated plasma from late pregnancy and could contribute in vivo to the higher reactivity of factor VII in this condition.