RESUMO
OBJECTIVE: The objectives were (1) to develop a cellular model of simulated ischaemia and reperfusion in isolated ventricular myocytes; (2) to determine effects of simulated ischaemia and reperfusion on calcium current (ICa), transient inward current (ITI) and contraction; and (3) to determine whether pharmacological agents which alter intracellular sodium and calcium loading affect signs of calcium overload in reperfusion in this model. METHODS: Electrical activity was recorded with conventional and voltage clamp techniques. Cell shortening was measured with a video edge detector. Myocytes were equilibrated in Tyrode solution, exposed to simulated ischaemia (hypoxia, acidosis, lactate, hyperkalaemia, glucose-free) for 20 min, and reperfused with Tyrode solution. RESULTS: Ischaemia depolarised myocytes [-89(SEM 1) to -67(4) mV, p < 0.05], abbreviated action potential duration [APD90, 257(14) to 188(12) ms, p < 0.05], and abolished contractions. Contractions elicited by voltage clamp steps also were abolished in ischaemia; however, ICa decreased by only 51% [-0.98(0.08) to -0.50(0.06) nA, p < 0.05]. Signs of calcium overload, including aftercontractions, oscillatory afterpotentials, and ITI, occurred in 69% of myocytes in reperfusion. Upon reperfusion, both APD90 and ICa recovered slowly; however, contractions returned quickly and temporarily exceeded control. Amiloride during ischaemia and reperfusion lowered incidence of ITI in reperfusion, whereas nifedipine and lignocaine had no effect on ITI. CONCLUSIONS: This model of ischaemia and reperfusion in ventricular myocytes shows many features of multicellular preparations, such as membrane depolarisation and action potential duration shortening during ischaemia, and appearance of oscillatory afterpotentials upon reperfusion. Inhibition of contraction during ischaemia and recovery of contraction in reperfusion are independent of changes in APD90 or ICa. Induction of aftercontractions, oscillatory afterpotentials, and ITI in reperfusion is associated with reduced peak ICa. Amiloride most probably decreased signs of calcium overload in early reperfusion by inhibiting sodium loading via Na+/H+ exchange. Additionally, amiloride may inhibit ITI directly by blocking Na+/Ca2+ exchange.
Assuntos
Modelos Animais de Doenças , Isquemia Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , Potenciais de Ação , Amilorida/farmacologia , Animais , ATPases Transportadoras de Cálcio/efeitos dos fármacos , ATPases Transportadoras de Cálcio/metabolismo , Tamanho Celular , Células Cultivadas , Cobaias , Coração/fisiopatologia , Masculino , Isquemia Miocárdica/fisiopatologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/metabolismoRESUMO
Acridine orange staining reveals changes within 3 hours of in vitro stimulation of normal rat lymphocytes with mitogens, and of immune rat lymphocytes with the sensitizing antigen. An increased number of red fluorescent cytoplasmic organelles, presumably lysosomes are seen by fluorescence microscopy. Fluorimetry of the supernatants from stained cell suspensions suggests an overall decreased cell uptake of the dye. The microscopy and fluorimetry detected early events in the reaction of lymphocytes from tumour-bearing rats with the target tumour cells. It would appear that the changes in intracellular behaviour of the dye and in overall cell uptake after immune stimulation are a reflection of dissociated variations in internal and external cell membrane permeability, and may provide simple general means for recognizing cellular immune reactions.
Assuntos
Acridinas , Memória Imunológica , Ativação Linfocitária , Linfócitos/imunologia , Acridinas/metabolismo , Animais , Antígenos de Neoplasias , Lectinas , Lisossomos/metabolismo , Microscopia de Fluorescência , RatosRESUMO
We examined the hypothesis that endogenous prostaglandins participate in the arrhythmogenic influence of ouabain in guinea pig hearts. Addition of ouabain (10 ng/ml) resulted in a 5-fold increase in the release of 6-keto-prostaglandin F1 alpha in the coronary effluent. Ten of 13 hearts studied (77%) demonstrated arrhythmic activity with a mean time to the onset of arrhythmias of approximately 35 min. The nonsteroidal antiinflammatory drugs indomethacin and acetylsalicylic acid which significantly inhibited the efflux of 6-keto-prostaglandin F1 alpha also reduced the incidence of arrhythmias to 10 of 30 hearts studied. In those hearts in which arrhythmias occurred, the time to onset was significantly increased to approximately 50 and 55 min for acetylsalicylic acid and indomethacin, respectively. In contrast, exogenous prostaglandin F2 alpha (0.1 and 1 ng/ml) and prostacyclin (0.1 and 10 ng/ml) increased the incidence of arrhythmias to 100% (10 of 10 hearts studied) and decreased the time to onset to approximately 10 min. These prostaglandin pretreatments were also able to reverse the protective actions of both acetylsalicylic acid and indomethacin. Other concentrations (10 ng/ml prostaglandin F2 alpha and 1 ng/ml prostacyclin) had no influence either on the incidence of arrhythmias or their time to onset. Prostaglandin E2 (0.1 ng/ml) produced a modest but not significant decrease in the time to onset of arrhythmias although this concentration was significantly effective in reversing the nonsteroidal antiinflammatory drug effects. The inotropic, chronotropic and coronary constricting actions of ouabain were unaffected either by nonsteroidal antiinflammatory drug or prostaglandin pretreatment. These studies suggest that prostaglandins are involved, at least in part, in the arrhythmogenic actions of ouabain in the isolated guinea pig heart.
Assuntos
Arritmias Cardíacas/fisiopatologia , Coração/efeitos dos fármacos , Ouabaína/toxicidade , Prostaglandinas/fisiologia , 6-Cetoprostaglandina F1 alfa/metabolismo , Animais , Arritmias Cardíacas/induzido quimicamente , Circulação Coronária/efeitos dos fármacos , Dinoprosta , Epoprostenol/farmacologia , Cobaias , Frequência Cardíaca/efeitos dos fármacos , Técnicas In Vitro , Masculino , Contração Miocárdica/efeitos dos fármacos , Prostaglandinas F/farmacologia , RadioimunoensaioRESUMO
Direct fluorescent antibody tests were used to detect Escherichia coli possessing K88 and 987P antigens. Identification of bacteria was accomplished on suspensions of organisms from clinical isolates, on frozen sections and impression smears from small intestine and on faecal smears. This assay makes possible the rapid identification of E. coli possessing K88 and 987P pilus antigens.
Assuntos
Adesinas de Escherichia coli , Antígenos de Bactérias/análise , Antígenos de Superfície/análise , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli , Escherichia coli/imunologia , Proteínas de Fímbrias , Fímbrias Bacterianas/imunologia , Doenças dos Suínos/microbiologia , Animais , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Fímbrias Bacterianas/ultraestrutura , Imunofluorescência , SuínosRESUMO
Enzyme-linked immunosorbent assay (ELISA) and crossed immunoelectrophoresis (IEP) were used to investigate antibody responses of sheep vaccinated with a double adjuvanted or single adjuvanted commercial foot rot vaccine. ELISA detected an antibody response of greater magnitude to the double adjuvant vaccine compared with the single adjuvant vaccine. Sera from sheep vaccinated with double adjuvant vaccine recognised at least six antigens of Bacteroides nodosus in crossed IEP while sera from the single adjuvant vaccinated sheep recognised one antigen. The use of non-denatured antigens of B nodosus in ELISA and crossed IEP enabled quantitative comparisons of antibody responses to the different foot rot vaccines to be made.
Assuntos
Anticorpos Antibacterianos/análise , Vacinas Bacterianas , Infecções por Bacteroides/veterinária , Bacteroides/imunologia , Pododermatite Necrótica dos Ovinos/prevenção & controle , Doenças dos Ovinos/prevenção & controle , Ovinos/imunologia , Animais , Infecções por Bacteroides/prevenção & controle , Ensaio de Imunoadsorção Enzimática , Imunoeletroforese Bidimensional , Masculino , Doenças dos Ovinos/microbiologiaRESUMO
Specific antibody to Eperythrozoon ovis was detected by an enzyme-linked immunosorbent assay (ELISA) in the sera of infected sheep. In the presence of parasite antigen, positive control serum showed a reaction approximately eight times that of negative serum. When compared to an immunofluorescent antibody test (IFAT), the ELISA was eight times more sensitive. Positive control sera gave a titre of 1:3200 by IFAT and 1:25,600 by ELISA. Through the use of a reference titration curve ELISA could be used as a semi-quantitative system to determine antibody levels in test sera.
Assuntos
Infecções por Anaplasmataceae/veterinária , Anticorpos Antibacterianos/análise , Infecções por Mycoplasma/veterinária , Mycoplasma/imunologia , Doenças dos Ovinos/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Infecções por Mycoplasma/imunologia , Ovinos , Doenças dos Ovinos/microbiologiaRESUMO
The effect of genotype on eating quality was evaluated on m. Longissimus thoracis et lumborum (LTL) muscle of 60 lambs. The lambs were sired by a selection of Texel (T), Poll Dorset (PD), Border Leicester (BL) and Merino (M) rams, crossed with Border Leicester x Merino (BLM) and Merino (M) ewes giving six genotypes (TxBLM, PDxBLM, TxM, PDxM, BLxM and MxM). The relationships between sensory panel assessment of eating quality attributes and pH, cooking loss and shear force were also investigated. No significant differences were observed between genotypes for panel assessment of tenderness, juiciness, aroma liking, aroma strength, flavour liking, overall acceptability and rating. MxM lambs had a significantly (P<0.05) higher flavour strength than BLxM lambs. pH was a poor indicator of any eating quality attributes, except aroma strength (r=0.3, P<0.05). Warner Bratzler shear force value (WB) and tenderness showed a significant (P<0.001) negative correlation (-0.7). Tenderness, flavour and juiciness were the most important sensory attributes, explaining 86.5% of the total variation in overall acceptability.
RESUMO
Successful protection against lumpy jaw disease in a colony of captive wallabies (Macropus eugenii) was induced by vaccination with a commercial ovine footrot vaccine. No mortalities attributable to lumpy jaw were observed in 69 vaccinated animals while six of 42 unvaccinated control wallabies died of the disease. Vaccinated animals exhibited significant increases in antibody titres to Bacteroides nodosus after the first and second doses of vaccine. Titres were measured by an enzyme-linked immunosorbent assay.
Assuntos
Vacinas Bacterianas/uso terapêutico , Infecções por Bacteroides/veterinária , Doenças Maxilomandibulares/veterinária , Macropodidae/imunologia , Marsupiais/imunologia , Animais , Anticorpos Antibacterianos/análise , Infecções por Bacteroides/prevenção & controle , Ensaio de Imunoadsorção Enzimática , Doenças Maxilomandibulares/microbiologia , Doenças Maxilomandibulares/prevenção & controleAssuntos
Antraz/veterinária , Bacillus anthracis/imunologia , Vacinas Bacterianas/imunologia , Doenças dos Bovinos/imunologia , Vacinação/veterinária , Animais , Antraz/epidemiologia , Antraz/imunologia , Antraz/prevenção & controle , Anticorpos Antibacterianos/biossíntese , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/prevenção & controle , Surtos de Doenças/prevenção & controle , Surtos de Doenças/veterinária , Feminino , Imunização Secundária/veterinária , New South Wales/epidemiologiaAssuntos
Infecções por Anaplasmataceae/veterinária , Eritrócitos/microbiologia , Infecções por Mycoplasma/veterinária , Mycoplasma/isolamento & purificação , Doenças dos Ovinos/microbiologia , Animais , Infecções por Mycoplasma/sangue , Infecções por Mycoplasma/microbiologia , Ovinos , Doenças dos Ovinos/sangueRESUMO
Transmembrane potentials and contractile activity were recorded from isolated canine Purkinje and ventricular muscle preparations exposed to acetylstrophantidin (AS) and subjected to a resting tension equal to 80% of that required to elicit peak developed tension. AS induced transient depolarizations (TD's) accompanied by aftercontractions in Purkinje tissue. AS also induced aftercontractions in 11 muscle preparations, and in seven of these the mechanical events were associated with TD's. Aftercontractions and TD's in both and the coupling intervals were directly related to the preceding basic cycle length (BCL). The amplitudes of aftercontractions and TD's reached a maximum at a coupling interval of 600-700 msec. Tension increased the amplitude of TD's in Purkinje tissue and promoted the appearance of TD's in muscle. TD's in muscle occasionally reached threshold in the presence of tension. The results of this study suggest that stretch or increased resting tension may promote the types of cardiac arrhythmias that are causally related to digitalis-induced TD's. The results are compatible with the hypothesis that TD's are caused either by a transmembrane influx of calcium or by an internal release of calcium ions.
Assuntos
Cálcio/fisiologia , Sistema de Condução Cardíaco/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Contração Miocárdica/efeitos dos fármacos , Ramos Subendocárdicos/efeitos dos fármacos , Estrofantidina/farmacologia , Estrofantinas/farmacologia , Animais , Cálcio/metabolismo , Cães , Ventrículos do Coração/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Miocárdio/metabolismo , Ramos Subendocárdicos/metabolismo , Estrofantidina/análogos & derivadosRESUMO
Digitalis-induced OAP provide a mechanism of automaticity that may be responsible for many arrhythmias induced by cardiac glycosides. In response to digitalis, OAP occur in tissues of the specialized conducting systems of both ventricles and atria and, under the influence of tension, occasionally in ventricular myocardium. Digitalis, in toxic doses, suppresses "normal" pacemaker activity possibly in part by enhancing overdrive suppression. In contrast to "normal" pacemaker activity, OAP exhibit, both in magnitude and rate of depolarization, postpacing acceleration. This plus the coupled nature of OAP are important characteristics in the generation of complex arrhythmias by OAP. Conduction disturbances may also be related to OAP. At early stages of intoxication OAP may speed conduction of superimposed beats relative to earlier or later beats. More advanced stages of intoxication are associated with conduction block. The occurrence of digitalis-induced OAP is promoted by high concentrations of calcium, low concentrations of potassium, and moderate stretch. OAP can be suppressed by high concentrations of potassium, reduction of extracellular calcium, and exposure to antiarrhythmic agents including diphenylhydantoin, verapamil, and aprindine. The effectiveness of the latter two agents may be related to ability to block transmembrane calcium currents. Digitalis-induced OAP in atrial tissue can be abolished by acetylcholine. A transmembrane current possibly but not necessarily carried by calcium appears to underly the occurrence of OAP. This current demonstrates kinetic properties different from those of the slow inward current associated with the plateau of the cardiac action potential. Calcium is intimately involved in the mechanism causing OAP and may be responsible for aftercontractions observed in conjunction with OAP. Aftercontractions greatly affect contractility and may be responsible at least in part for some of the inotropic actions of digitalis. Thus the occurrence of OAP may be linked to the inotropic actions of digitalis. Digitalis-induced OAP provide a mechanism of automaticity with characteristics paralleling automatic behavior observed in intact animals intoxicated with digitalis. The relative importance of OAP in the genesis of clinically important arrhythmias awaits further investigation.
Assuntos
Arritmias Cardíacas/induzido quimicamente , Glicosídeos Digitálicos/efeitos adversos , Potenciais de Ação/efeitos dos fármacos , Animais , Antiarrítmicos/farmacologia , Arritmias Cardíacas/fisiopatologia , Cálcio/fisiologia , Complexos Cardíacos Prematuros/fisiopatologia , Glicosídeos Digitálicos/farmacologia , Estimulação Elétrica , Átrios do Coração/fisiopatologia , Bloqueio Cardíaco/fisiopatologia , Sistema de Condução Cardíaco/fisiopatologia , Frequência Cardíaca , Ventrículos do Coração/fisiopatologia , Humanos , Íons , Potenciais da Membrana , Contração Miocárdica/efeitos dos fármacos , Potássio/farmacologia , Potássio/fisiologia , Ramos Subendocárdicos/fisiopatologia , Estrofantidina/análogos & derivados , Estrofantidina/farmacologia , Nervo Vago/fisiopatologiaRESUMO
Contractile activity was recorded from isolated canine ventricular muscle exposed to acetylstrophanthidin (AS), 0.5 to 2 X 10(-7) g/ml. Development of the positive inotropic effect of AS was accompanied by the appearance of aftercontractions (AC) coupled to the driven responses. The amplitudes of AC increased with the number of preceding beats. Test beats occurring during the ascending limb of AC were potentiated and the potentiation increased with the amplitude of the AC. Beats falling during the descending limb decreased in strength as the amplitude of the AC increased. As eliminated the frequency dependence of restitution. However, staircase phenomena persisted and were then found to be dependent on the phase relationship of each beat to underlying AC. These findings suggest an important role for AC in the inotropic actions of digitalis and in determining strengh-interval relationships of ventricular muscle treated with digitalis.
Assuntos
Contração Miocárdica/efeitos dos fármacos , Estrofantidina/análogos & derivados , Animais , Cães , Feminino , Sistema de Condução Cardíaco/efeitos dos fármacos , Frequência Cardíaca , Ventrículos do Coração/efeitos dos fármacos , Masculino , Estrofantidina/farmacologiaRESUMO
Transmembrane activity was recorded from isolated canine false tendons or trabeculae by using standard differential microelectrode techniques. Exposure of the tissues to acetylstrophanthidin (AS; 0.5-2 X 10(-7) g/ml) induced oscillatory afterpotentials in false tendons but not in muscle. When the transmembrane potential of the isolated tissues was altered by externally applied current, progressive depolarization of false tendons caused the amplitude of oscillatory afterpotentials to increase to a maximum and then decrease. Hyperpolarization abolished oscillatory afterpotentials and unmasked activity attributable to the normal pacemaker mechanism. Any level of manifest oscillatory afterpotential-related toxicity, including phasic decreases in excitability, could be elicited by careful selection of membrane potential. The effects of the imposed membrane potentials were immediately reversed with the return of membrane potential to control levels. In muscle, exposed to AS but not exhibiting oscillatory afterpotentials, depolarization revealed oscillatory afterpotentials. In both false tendons and muscle, depolarization to membrane potentials of -50 mV or less before exposure to AS elicited depolarization-induced automaticity. Subsequent exposure of the tissues to AS abolished this activity. This study demonstrates an important role of membrane potential in digitalis toxicity.
Assuntos
Coração/efeitos dos fármacos , Estrofantidina/análogos & derivados , Animais , Cães , Coração/fisiologia , Sistema de Condução Cardíaco/efeitos dos fármacos , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Ramos Subendocárdicos/efeitos dos fármacos , Ramos Subendocárdicos/fisiologia , Estrofantidina/farmacologiaRESUMO
Electrical activity from endo- and epicardium was recorded from isolated segments of right ventricular free walls of guinea pigs using standard differential microelectrode techniques and a high-gain electrocardiogram (ECG). Stimulation was applied to the endocardium. Tissues were exposed to ischemic conditions for 10 min and then were reperfused with "normal" Tyrode's solution. Early premature beats or rapid ventricular tachycardia (VT) occurred in 36% of hearts during "ischemia" and 79% of hearts on reperfusion. Endocardial activation was not significantly slowed by ischemic conditions or reperfusion. However, transmural conduction times increased, and muscle action potential durations decreased during ischemic conditions and early reperfusion. Rapid VT began with alternating activation of endo- and epicardium, and continuous low-voltage ECG activity bridging diastole. Activation of epicardium was essential for occurrence of early premature beats and rapid VT. Hyperkalemia during ischemia promoted arrhythmias during "ischemia," but not during reperfusion. Oscillatory afterpotentials (OAP) also occurred during reperfusion (36%), but not during "ischemia". Our study provides an isolated tissue model that reproducibly generates tachycardias, and that permits study of ischemia and reperfusion-induced transmembrane activity and defects in transmural conduction.
Assuntos
Doença das Coronárias/fisiopatologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Taquicardia/fisiopatologia , Potenciais de Ação/fisiologia , Animais , Doença das Coronárias/complicações , Estimulação Elétrica , Endocárdio/fisiopatologia , Cobaias , Sistema de Condução Cardíaco/fisiopatologia , Ventrículos do Coração/fisiopatologia , Técnicas In Vitro , Masculino , Ramos Subendocárdicos/fisiopatologia , Taquicardia/etiologia , Função Ventricular Direita/fisiologiaRESUMO
Transmembrane electrical activity from endo- and epicardium and a high-gain ECG were recorded from isolated segments of guinea pig right ventricles. Endocardium was stimulated. Tissues were exposed to ischemic conditions for 15 min and then reperfused with "normal" Tyrode's solution. Ventricular tachycardia, bigeminy, or trigeminy with characteristics of transmural reentry occurred in early reperfusion in 68% of control hearts. Arrhythmias were associated with prolongation of the transmural conduction time (CT) and abbreviation of the endocardial effective refractory period (EP). Quinidine significantly suppressed reperfusion arrhythmias at 1 and 5 microM, slightly increased the incidence of arrhythmias at 10 microM, and again suppressed arrhythmias at 50 and 100 microM. At 1 and 5 microM, quinidine prevented or attenuated prolongation of the transmural CT by ischemic conditions and reperfusion. The transmural CT was not significantly changed at 10 microM, and was further prolonged at 50 and 100 microM quinidine. The endocardial ERP was prolonged by 50 and 100 microM quinidine during ischemic conditions and reperfusion. In epicardial slices, 5 microM quinidine shortened the CT transverse to the fiber orientation during reperfusion but had no effect on the longitudinal CT. Thus, antiarrhythmic efficacy of low concentrations of quinidine may occur through differential effects dependent on tissue anisotropy.
Assuntos
Arritmias Cardíacas/fisiopatologia , Doença das Coronárias/fisiopatologia , Sistema de Condução Cardíaco/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Quinidina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Arritmias Cardíacas/etiologia , Doença das Coronárias/complicações , Eletrofisiologia , Cobaias , Técnicas In Vitro , Função Ventricular Direita/efeitos dos fármacosRESUMO
The effects of verapamil and nifedipine on cellular mechanisms of arrhythmia were examined in isolated canine Purkinje fiber-papillary muscles. Microelectrode recordings were made simultaneously from both tissues. Preparations were superfused with Tyrode's solution modified to mimic specific conditions of ischemia for 40 min with or without calcium channel blockers. Verapamil or nifedipine resulted in significantly greater depolarization of Purkinje tissue in response to ischemic conditions and increased the incidence of inexcitability or conduction block in Purkinje and muscle tissues. These calcium channel blockers caused only minor changes in ischemia-induced depolarization of muscle. In Purkinje tissue, return to nonischemic conditions in the absence of drugs caused, in sequence, oscillatory afterpotentials, temporary depolarization to inexcitability, and a phase of automaticity at low membrane potential. These events did not occur in muscle. Verapamil or nifedipine abolished oscillatory afterpotentials and low membrane potential automaticity in Purkinje tissue. However, reperfusion-induced depolarization and inexcitability of Purkinje tissue was delayed but not attenuated. This study demonstrates that verapamil or nifedipine exacerbate depolarization and depression of conduction in Purkinje tissue exposed to ischemic conditions. However, verapamil and nifedipine suppress some but not all potential mechanisms of arrhythmia induced by reperfusion.
Assuntos
Arritmias Cardíacas/tratamento farmacológico , Nifedipino/farmacologia , Traumatismo por Reperfusão/tratamento farmacológico , Verapamil/farmacologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Diástole/efeitos dos fármacos , Diástole/fisiologia , Cães , Eletrofisiologia , Sistema de Condução Cardíaco/efeitos dos fármacos , Sistema de Condução Cardíaco/fisiologia , Masculino , Potenciais da Membrana/efeitos dos fármacos , Ramos Subendocárdicos/efeitos dos fármacos , Ramos Subendocárdicos/fisiologia , Traumatismo por Reperfusão/fisiopatologiaRESUMO
Catecholamines increase the amplitudes of oscillatory afterpotentials (OAP) and peak magnitude of the transient inward current (Iti) responsible for OAP. The objectives of this study were to determine whether beta-adrenoceptor stimulation can induce Iti, and to determine the mechanism by which beta-adrenoceptor stimulation increases the magnitude of Iti. Experiments were performed using standard two electrode voltage--clamp techniques in isolated rabbit Purkinje fibers. Holding potential was either -50 or -80 mV. The Iti was elicited by repolarizing steps, following 1.5 or 3 s activating steps to potentials near 0 mV. Isoproterenol (ISO) failed to induce the Iti at concentrations from 10(-8) to 10(-6)M. However ISO (10(-7)M) significantly increased peak magnitude of spontaneously occurring Iti (P less than 0.05), or Iti induced by acetylstrophanthidin (AS) (P less than 0.05). ISO also shifted the minimum activation voltage 10 mV more negative (P less than 0.05). The current-voltage relationship demonstrated that ISO significantly increased the range of potentials over which Iti greater than or equal to 5 nA occurred, but did not significantly shift the voltage at which maximum peak current was observed. Effects of ISO on Iti were blocked by 10(-7)M propranolol or atenolol. Mn2+ (2 mM) or verapamil (2 microM) blocked the slow inward current (Isi) more than 80% before substantially decreasing peak Iti. Either agent blocked stimulation of Isi but not Iti by ISO at 10(-7)M. In contrast, quinacrine (20 microM), an inhibitor of Na(+)-Ca2+ exchange, abolished stimulation of Iti by ISO while having no significant effect on Isi. Our results indicate that beta-adrenoceptor stimulation cannot induce Iti in rabbit Purkinje fibers, but can enhance the Iti induced by other means, by stimulating Na(+)-Ca2+ exchange.
Assuntos
Arritmias Cardíacas/metabolismo , Isoproterenol/farmacologia , Ramos Subendocárdicos/metabolismo , Receptores Adrenérgicos beta/metabolismo , Animais , Cálcio/metabolismo , Técnicas In Vitro , Troca Iônica , Cinética , Manganês/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Ramos Subendocárdicos/efeitos dos fármacos , Quinacrina/farmacologia , Coelhos , Receptores Adrenérgicos beta/efeitos dos fármacos , Sódio/metabolismoRESUMO
Transmembrane electrical activity was recorded from endo- and epicardium of isolated segments of guinea pig right ventricles with standard microelectrode techniques. An ECG was also recorded by two electrodes placed at opposite ends of the tissue bath. Regular stimulation was delivered to the endocardium. Tissues were exposed to simulated ischemia for 15 min and then were reperfused with "normal" Tyrode's solution. Rapid sustained or nonsustained ventricular tachycardia, bigeminy or trigeminy with characteristics of transmural reentry occurred in early reperfusion in 14 of 20 hearts (70%). Arrhythmias were accompanied by prolongation of transmural conduction time and abbreviation of endocardial effective refractory period. With lidocaine, at 1, 5, 10 and 50 microM, reperfusion arrhythmias occurred in 53.3, 22.2, 20.8 and 14.3% of hearts, respectively. The decreased incidence of arrhythmias was statistically significant for 5 to 50 microM lidocaine (P less than .01). The antiarrhythmic effect did not correlate with changes in transmural conduction time, endocardial effective refractory period, or endocardial excitability. However, antiarrhythmic concentrations of lidocaine selectively depressed epicardial excitability and significantly increased endo- to epicardial conduction block during late ischemic and early reperfusion periods. Epicardial inexcitability extended to late diastole and conduction block was not restricted to premature beats. Thus, in transmural reentry in which the epicardium is an essential component of the circuit, lidocaine may interrupt the circuit by selectively rendering this component inexcitable.
Assuntos
Arritmias Cardíacas/prevenção & controle , Lidocaína/farmacologia , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Potenciais de Ação/efeitos dos fármacos , Animais , Arritmias Cardíacas/fisiopatologia , Diástole/efeitos dos fármacos , Eletrofisiologia , Cobaias , Bloqueio Cardíaco/fisiopatologia , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/fisiopatologia , Técnicas In Vitro , Masculino , Modelos Cardiovasculares , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Oxigênio/fisiologiaRESUMO
1. Membrane currents were measured with a two-microelectrode technique in voltage clamped rabbit cardiac Purkinje fibres under conditions known to cause intracellular calcium overload and to eliminate or minimize Na(+)-Ca2+ exchange. 2. Increasing [Ca2+]o from 2.5 to 5 mM or above and substituting external sodium with either sucrose, choline or Li+ induced an oscillatory transient inward current (TI) which peaked 200-300 ms after repolarization from a previous depolarizing pulse. The TI quickly disappeared upon return to normal Tyrode solution. Both the rate and configuration of action potentials of Purkinje fibres also returned to control upon return to Tyrode solution after 30 min of high Ca2+ exposure, if the Ca2+ concentration was 30 mM or less. 3. The TI in Na(+)-free solution was Ca2+ dependent. Either zero or low (2.5 mM) [Ca2+]o, or replacement of [Ca2+]o by BaCl prevented induction of the TI current upon repolarization from a previous depolarizing pulse. 4. In the presence of 30 mM-CaCl2 and with choline chloride as the substitute for NaCl, TI had a distinct reversal potential (Erev) of -25 mV. The time-to-peak TI, either inward or outward, did not shift significantly with change in voltage. Both inward and outward TI were simultaneously abolished by exposure to 1 microM-ryanodine, suggesting they were both activated by transient release of Ca2+ from the sarcoplasmic reticulum. The occurrence of TI in the absence of [Na+]o is not compatible with an electrogenic Na(+)-Ca2+ exchange mechanism. The existence of a clear-cut reversal potential suggests that an ionic channel may be responsible for the TI under these conditions. 5. Both the magnitude of peak TI and the Erev were affected by changes of CaCl2 concentration. (i) Under steady-state conditions, peak inward TI was significantly increased when the [Ca2+]o was elevated from 5 to 15 mM. The peak TI in the outward direction was significantly increased when [Ca2+]o was elevated from 15 to 30 mM; however, the difference in peak inward TI at 15 and 30 mM [Ca2+]o was small. (ii) Clear-cut reversals of TI were found at Ca2+ concentrations of 10 mM (Erev = -19.5 mV) or greater, and elevation of [Ca2+]o to 20, 30, 50 and 105 mM shifted the Erev to more negative potentials. (iii) In the presence of 5 mM [Ca2+]o the inward TI declined to zero at about -30 mV, and test voltages between -55 and +5 mV failed to reveal a distinct outward TI.(ABSTRACT TRUNCATED AT 400 WORDS)