Acetaldehyde depresses myocardial contraction and cardiac myocyte shortening in spontaneously hypertensive rats: role of intracellular Ca2+.

Acetaldehyde (ACA), the major metabolite of ethanol, exerts both stimulatory and depressive actions on myocardial tissue. We have recently shown that ACA depresses myocardial contraction, cardiac myocyte shortening and intracellular Ca2+ transients in normal rat heart. The purpose of the present study was to determine the influence of hypertension on ACA-induced myocardial actions. Mechanical properties of left ventricular papillary muscles and ventricular myocytes isolated from both 25-week-old normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) were evaluated using force-transducer and video edge-detection, respectively. Papillary muscles and cardiac myocytes were electrically stimulated to contract at 0.5 Hz. Contractile properties analyzed include: peak tension development (PTD), peak twitch amplitude (PTA), time-to-PTD/PTA (TPT/TPS), time-to-90% relaxation/relengthening (RT90/TR90) and maximal velocities of contraction/shortening and relaxation/relengthening (+/-VT/+/-dL/dt). Intracellular Ca2+ transients were measured as fura-2 fluorescence intensity (FFI) changes. ACA (1-30 mM) depressed PTD without affecting other mechanical indices in both WKY and SHR myocardium, with maximal inhibition of 64 and 69%, respectively. SHR myocytes exhibited increased cell dimension, baseline PTA and resting intracellular Ca2+ levels, compared to WKY counterparts. ACA (0.03-30 mM) depressed PTA without affecting TPT, TR90 and +/-dL/dt. The maximal inhibitions were 31 and 36% in WKY and SHR groups, respectively. Interestingly, ACA exerted a biphasic effect on FFI, displaying potentiation at lower doses (<3 mM) and inhibition at higher doses (>3 mM). The maximal increase in FFI changes were 19 and 22% at 0.3 mM and the maximal decreases were 37 and 29% at 30 mM ACA, in WKY and SHR myocytes, respectively. Neither resting intracellular Ca2+ levels (FFI) nor fluorescence decay time (FDT) were affected by ACA. The increase in FFI was attenuated by propranolol (1 microM), whereas the decrease in FFI was reversed by BayK 8644 (1 microM). These results suggest that hypertension does not appear to alter ACA-induced myocardial depression. The mechanism underlying ACA-induced myocardial actions may involve increased beta-adrenergic activity at low doses and reduced Ca2+ entry and/or release at high doses.

Influence of age on the inotropic response to acute ethanol exposure in spontaneously hypertensive rats.

Acute ethanol exposure depresses cardiac electromechanical function, whereas chronic ethanol consumption leads to the development of a specific myopathic state. Chronic hypertension and aging have similar effects in the impairment of myocardial function. However, little is known about the effects of ethanol on cardiac mechanical function in hypertension. We studied the effect of age on baseline mechanical properties and the inotropic response to clinically relevant concentrations of ethanol (18 to 71 mmol/L) using papillary muscles from spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) at 10 and 25 weeks of age. Mechanical parameters measured were peak tension developed, time to peak tension, time to 90% relaxation, and maximal velocities of tension development and tension decline. SHR exhibited elevated systolic pressure and body weight as well as cardiomegaly and hepatomegaly at 10 and 25 weeks of age. Baseline mechanical properties were similar in SHR and WKY muscles at 10 weeks, whereas at 25 weeks, SHR muscles developed less tension, and both maximal velocities of tension development and tension decline were markedly depressed. Ethanol exposure produced concentration-dependent negative inotropic effects in both groups at both ages. Ethanol (> 18 nmol/L) decreased peak tension developed in both groups at 10 weeks, although higher concentrations were required at 25 weeks. The negative inotropic effect of ethanol resulted in the shortening of time to 90% relaxation in both groups at 10 weeks and was associated with a slowing of maximal velocities of both tension development and tension decline. The results suggest that aging depresses baseline mechanical properties when coupled with hypertension. In addition, the magnitude of the negative inotropic effect of ethanol was attenuated in both groups at 25 weeks of age.

Acute and chronic effects of ethanol on papillary muscles from spontaneously hypertensive rats.

The effects of chronic ethanol ingestion (12 weeks) on the mechanical properties of hypertrophied papillary muscle and the in vitro effects of ethanol (80-640 mg/dl) was studied. Papillary muscles from spontaneously hypertensive rats (SHRs) and their normotensive controls, the Wistar-Kyoto rat (WKY), were used in this study. Peak-developed tension was significantly less in muscles obtained from SHR compared with WKY even when normalized for muscle cross-sectional area. Chronic ethanol ingestion resulted in a significant shortening of both contraction and relaxation duration in muscles from SHR and WKY. In muscles from SHR and WKY, acute in vitro ethanol exposure produced concentration-dependent negative inotropic effects that were associated with a reduction in the duration of contraction and relaxation and marked slowing in the maximum velocities of tension development and decay. These findings suggest that the contractile response to ethanol exposure, in vitro, is not modified by either chronic ethanol ingestion or hypertension.

Effect of capillary pressure and lung distension on capillary recruitment.

To investigate the effect of capillary pressure and alveolar distension on capillary recruitment, we used video-microscopy to quantify capillary recruitment in individual subpleural alveolar walls. Canine lobes were perfused with autologous blood either while inflated by positive airway pressure or while inflated by negative intrapleural pressure in the intact thorax with airway pressure remaining atmospheric. Low flow rates minimized the arteriovenous pressure gradient (< 5 mmHg), permitting capillary pressure estimation by averaging these pressures. Capillary pressure was varied stepwise from airway pressure to 30 mmHg above airway pressure. Capillary recruitment always began as capillary pressure exceeded airway pressure. At low positive airway pressures, the capillaries of the excised lobes opened suddenly over a narrow pressure range. AT higher airway pressures and in the intact thorax, recruitment occurred over a wide range of capillary pressures. We conclude that capillary perfusion begins when intracapillary pressure just exceeds alveolar pressure but that further increases in capillary pressure recruit capillaries depending on tension in the alveolar wall, whether imposed by positive airway pressure or by gravity when the lung is suspended in an intact thorax.

Effect of increasing flow on distribution of pulmonary capillary transit times.

The complex morphology of the pulmonary capillary network causes capillary transit times to be dispersed about a mean. It is known that flow-induced decreases in mean capillary transit time are partially offset by capillary recruitment and distension, but the effect of these factors on the rest of the distribution of transit times is unknown. We have studied the relationship between blood flow, capillary recruitment, and the distribution of transit times in isolated canine lungs with videomicroscopy. Doubling baseline lobar blood flow recruited capillaries. All transit times in the distribution decreased, as did relative dispersion. Doubling flow again caused a further decrease in transit times, but neither capillary recruitment nor relative dispersion changed significantly. We conclude that capillary transit times become more homogeneous as lobar flow increases from low to intermediate levels. Further increases in flow across a fully recruited network are associated with decreases in transit times but not with more homogeneous capillary perfusion.

Computer simulation of neutrophil transit through the pulmonary capillary bed.

One-half of the neutrophils that enter the pulmonary circulation become temporarily trapped in capillaries. The neutrophils that are impeded make complete stops between free-flowing movements. These observations, based on in vivo microscopy, suggest that pulmonary margination is caused by neutrophils being impeded at focal sites in the capillary bed. To investigate the frequency with which impeding sites had to occur in the pulmonary capillaries to trap one-half of the circulating neutrophils, we developed a computer model to simulate neutrophils encountering discrete obstructions in a capillary-like network. Surprisingly, if only 1% of the capillaries in the network acted as traps, one-half of the neutrophils stopped at least once. The trapping ability of a given percentage of obstructions was independent both of the geometry of the network was whether the obstructions occurred in the segments or junctions. To simulate neutrophil transit more realistically, both neutrophil and capillary diameters were randomly selected from published diameter distributions. Every neutrophil was trapped multiple times by this model, suggesting that cell deformation contributes importantly to neutrophil passage through the pulmonary capillary bed.

Effect of increased left atrial pressure on breathing frequency in anesthetized dog.

Distension or loading of the isolated canine left heart caused reflex tachypnea in prior studies. The object of the present effort was to explore the possibility that this depended primarily on atrial distension. Cardiopulmonary bypass perfusion and ligation of pulmonary veins were used to isolate the left-heart chambers of anesthetized dogs. Simultaneous distension of the beating left atrium and fibrillating ventricle stimulated breathing frequency (f), whereas isolated ventricular distension did not. At other times, intervals of atrial fibrillation were imposed under two different conditions: 1) while the right heart and lungs were bypassed and systemic perfusion was provided by the left ventricle using blood returned to the left atrium by pump and 2) while the ventricles fibrillated and systemic perfusion was supplied directly by the pump. Atrial fibrillation increased left atrial pressure and stimulated f in condition 1. In condition 2, f increased only if fibrillation was associated with a rise in left atrial pressure. Vagal cooling blocked the effect of fibrillation. I conclude that left atrial distension may initiate reflex tachypnea.

Effect on breathing of abruptly loading and unloading the canine left heart.

Cardiopulmonary bypass and pulmonary vein ligation were used to isolate left hearts of anesthetized open-chest dogs. After external gas exchange, blood was returned at constant flow (approximately 120 ml.min-1.kg-1) directly to the aorta or indirectly through the left heart ("left heart loading"). Loading caused breathing frequency (f) to increase approximately 5 breaths/min (approximately 20%), whereas systemic arterial pressure (Psa) fell approximately 15%. Because Psa was pulsatile during loading, we demonstrated separately the effect of pulsatile pressure and found it to lower mean Psa without changing f. Cooling cervical vagi to 7 degrees C eliminated the f response to loading and slightly decreased the Psa response. Loading was compared with graded distension of the fibrillating ventricle and beating atrium, which also increased f. As measured by an abdominal pneumograph, depth of breathing decreased significantly (approximately 4%) during left heart loading but did not change significantly on distension of the fibrillating heart. I conclude that left heart loading may induce tachypnea and a slightly reduced tidal volume by a vagal reflex most likely originating from the left heart.

Breathing response to lung congestion with and without left heart distension.

This study compared the effect of lung congestion with and without left heart (LH) distension on breathing frequency (fr) and discriminated among responses mediated by myelinated and nonmyelinated vagal afferents. Cardiopulmonary bypass perfusion of anesthetized dogs was used to isolate reflexes. The following three groups were prepared: 1) lung vessels pressurized by pumping into the main pulmonary artery (MPA); 2) lungs and fibrillating LH pressurized by pumping into MPA while draining from LH; 3) lungs congested by occluding several pulmonary veins while holding cardiac output constant. Congestion of lungs alone in groups 1 and 3 depressed fr. Congestion of lungs and distension of LH (group 2) caused transient depression of fr but a steady-state excitation. Cooling cervical vagi to 8 degrees C prevented depression of fr by congestion in all groups. In groups 1 and 2, in which MPA pressure was higher than in group 3, congestion during vagal cooling stimulated breathing. I conclude that lung congestion may stimulate fr via C-fiber afferents, but this may be overcome by a depressor effect via myelinated afferents. Simultaneous LH distension may reflexly stimulate breathing and overcome the lung depressor reflex.

Effects of lung congestion and oleic acid injury on the Hering-Breuer reflex.

Breathing and the Hering-Breuer (HB) reflex may be stimulated by congestion and by acute lung injury, but there is disagreement about the effects of both stimuli. This study evaluated these effects using greater stimulus isolation and control of secondary interactions than have previously been employed. Pressurization of lung vessels and left heart and oleic acid injury were individually imposed on anesthetized open-chest dogs perfused with an external pump and gas exchanger. Lungs were inflated in steps before and during those stimuli. The HB reflex was evaluated from graphs of breathing frequency (fr) vs. airway pressure. Congestion itself had no significant sustained effect on fr, but it slightly depressed the HB reflex. Oleic acid tachypnea that was depressed to pretreatment fr by inflation, implying enhancement of the HB response. Capsaicin and oleic acid had similar effects. Vagal cooling to 8 degrees C slightly depressed the effects of oleic acid and capsaicin, had no effect on the sustained fr response to congestion, and reversed the inhibitory effect of inflation. A stimulation of breathing or an enhancement of the HB reflex by congestion was not confirmed, but oleic acid increased fr and the HB reflex.

Control of breathing in anesthetized dogs by a left-heart baroreflex.

Anesthetized open-chest dogs on cardiopulmonary bypass were used to test the hypothesis that breathing reflexly responds to distension of the left-heart chambers. Bypass perfusion withdrew systemic flow from the right atrium and returned it to the aorta after gas exchange. Ventricles were fibrillated. The left heart was isolated by tying all pulmonary veins, and it was perfused separately at low flow admitted through one pulmonary vein and withdrawn from the ventricle. Left-heart pressure was intermittently raised abruptly from a nominal base line of 0 by partial occlusion of outflow. Pressures from approximately 10 to 50 cmH2O caused proportional increases in breathing frequency and decreases in expiratory and inspiratory times. Changes occurred immediately, reached a plateau within approximately 20 s, and were sustained for periods of observation as long as 3 min. Recovery to base line followed stimulus removal. Vagal cooling to 8 degrees C prevented responses, but autonomic ganglion blockade with hexamethonium had no effect. I conclude that breathing may be stimulated by left-heart distension and that this is mediated by large myelinated vagal afferents.

Pulmonary arterial distension does not cause pulmonary vasoconstriction.

Distension of the main pulmonary artery or its major branches with an intraluminal balloon has been reported to cause pulmonary vasoconstriction by an unknown mechanism. This study was an attempt to confirm the pressor response and explore its cause. Several balloon distension methods were tried and discarded because they caused unintentional obstruction. Ultimately, I inflated a balloon placed retrogradely and confined to the left main pulmonary artery of six anesthetized open-chest dogs after ligating left lobar arterial branches. Blood flow and systemic gas composition were controlled by interposing an external pump oxygenator between the left ventricle and aorta. Pressures in the aorta, main pulmonary artery, and left atrium were recorded. Alveolar hypoxia was used as an independent test of pulmonary vasoreactivity. Although hypoxic pressor responses occurred, challenges with arterial distension did not change lung perfusion pressure. Silicone rubber casts were made of the arteries of six dogs used in pilot experiments. These revealed the limited lengths in which distenders can be placed without unintentional encroachment on flow. I could not support the conclusion that arterial distension causes vasoconstriction and am suspicious that the perfusion pressure increases reported by others may have been caused by undetected obstruction of a major arterial branch.

Pulmonary hypertension induced with an intra-arterial balloon: an alternate mechanism.

The Laks catheter is a triple-lumen balloon catheter used to distend the canine main pulmonary artery while recording right ventricular pressure and the arterial pressure distal to the balloon. A rise in arterial pressure reported to occur during distension has been attributed to vasoconstriction rather than passive obstruction by the balloon. We tested this in six anesthetized dogs by inflating the Laks catheter-balloon while recording pressure distal to the balloon from the Laks catheter as well as from additional catheters in right and left pulmonary arteries placed retrogradely through lobar branches following thoracotomy. We found that balloon inflation increased pressures in the arterial port of the Laks catheter and in the left pulmonary artery catheter but reduced it in the right pulmonary artery. Tightening a snare around the right pulmonary artery had the same effects on pressures. Similar results were obtained while cardiac output was controlled by left ventricular bypass perfusion in four dogs. We conclude that the Laks catheter-balloon obstructs flow to the right lung and that the arterial pressure rise recorded in it during balloon inflation cannot be distinguished from that caused by occlusion of the right pulmonary artery.

Effect on breathing of acute pressure rise in pulmonary artery and right ventricle.

We tested the hypothesis that breathing would be regulated in response to right ventricular and pulmonary arterial pressure changes when secondary events are controlled. Dogs were anesthetized, thoracotomies were performed, and cardiopulmonary bypass perfusion was established. Lungs were inflated to sustained pressures. The left diaphragmatic lobe was retrogradely cannulated and all other lobar arteries were ligated, forming a pulmonary arterial sac that drained to the oxygenator from the cannula and filled from systemic venous return by the beating right ventricle. Right atrial pressure was adjusted to produce sac flows of approximately 400 ml/min. We recorded systemic and pulmonary arterial pressures, sac flow, and the integrated diaphragm electromyogram (DEMG). Resistive loads were imposed on sac outflow by adjusting a clamp. Loaded mean pulmonary arterial pressures ranged from 27 to 70 Torr. Loading increased respiratory frequency without affecting peak DEMG amplitude. Responses did not occur after vagotomy. Effects were quantitatively modest: pressurization to approximately 50 Torr increased frequency approximately 3.4 breaths/min (22%). The magnitude of change was insufficient to explain in intact dogs the ventilatory responses that have been attributed to this reflexogenic unit.

Effect of inspiration on inferior vena caval blood flow in dogs.

Inferior vena cava flow of anesthetized open-chest dogs was drained to a reservoir from a cannula above the diaphragm and returned to the atrium at constant rate. At selected base-line caval pressures, the caval flow and pressures in the abdomen (Pab), iliac vein (Piv), and downstream cavae (Pvc) were recorded during spontaneous breathing, cyclic phrenic nerve stimulation, and cyclic lowering of caval drain pressure. Each augmented flow unless Pab exceeded Pvc by at least ca. 5 cmH2O. In other dogs a cannulating flow probe was placed in the thoracic inferior cava and the chest was reclosed. Flow was augmented throughout most or all of spontaneous inspiration and was never depressed even though Pab exceeded right atrial pressure and Piv. I conclude that the collapse of hepatic veins and proximate cava does not occur at most normal pressures and a Starling resistor analog of abdominal veins based solely on abdominal and venous pressures is inappropriate. Both falling atrial pressure and rising Pab probably augment inspiratory abdominal venous return.

Effect of diaphragm contraction on canine heart and pericardium.

Pericardiophrenic attachments transmit diaphragm contraction to the pericardium. We investigated this in two ways. 1) We replaced the hearts of externally perfused dogs with a balloon from which we measured pressure changes. Diaphragm contraction increased pressure from 4.6 to 5.5 Torr, equivalent to an isobaric volume decrease of 1.5%, and decreased volumetric compliance by 3%. 2) We selectively servo controlled right atrial pressure, left atrial pressure, or cardiac output in open-chest dogs and monitored the effect of diaphragm contraction on cardiovascular and abdominal pressures, cardiac output, and the volume of blood added to or withdrawn from the circulation to achieve servo control. Diaphragm contraction decreased left atrial pressure 0.4 Torr when right atrial pressure was controlled and right atrial pressure increased 0.2 Torr while controlling left atrial pressure, but there were no significant changes in cardiac output. Atrial pressure did not change significantly when output was controlled. Servo control required removal of approximately 50 ml of blood, presumably reflecting a decreased splanchnic vascular capacity at the higher abdominal pressure. We conclude that the diaphragm may slightly tense the pericardium, but this has no important primary effect on the heart.

Respiratory system compliance as seen from the cardiac fossa.

Changes in cardiac size and shape should impose stresses on the surrounding lung and chest wall. To examine pressure-volume relationships of the cardiac fossa we measured pressures required to increase the pericardial volume of freshly killed dogs at different levels of lung inflation, first by expanding the pericardium uniformly and then by expanding only the left ventricle. In both cases we obtained linear pressure-volume relationships, the slopes of which expressed an apparent compliance. Compliance decreased as lung volumes were increased by raising end-expiratory pressure, and compliance with symmetrical pericardial filling exceeded that with asymmetrical (left ventricular) distension. These compliances were compared with the total respiratory system compliance measured during tidal ventilation, and we found that the compliance of the cardiac fossa was significantly less than would be predicted from lung and chest wall compliances as classically measured. We concluded that the respiratory system imposes a finite compliance load on cardiac filling that raises local epicardial pressure above ambient pleural pressure. This respiratory system load depends upon lung volume and the cardiac shape change.

Effect of oleic acid injury on lung-heart interaction during ventricular filling.

Part of ventricular filling pressure is expended in deforming the lungs and oleic acid lung injury, which reduces lung compliance, might be expected to change the magnitude of this effect. Left ventricular pressure-volume curves from 13 treated and 13 uninjured dogs were compared. Curves were obtained by inflating a balloon in the flaccid left ventricle of the freshly dead dog. The extracardiac compliance component was determined by subtracting ventricular pressures observed when lungs were collapsed from those observed during inflation to levels duplicating both airway pressure and lung volume of uninjured dogs. We found that the respiratory system compliance opposing ventricular filling is much less than is predicted from the compliance opposing tidal ventilation. This suggests that cardiac filling imposes a local shape change rather than a general lung volume change. Injury only slightly decreased cardiac filling compliance although it markedly reduced lung inflating compliance. These results are consistent with citations showing that edema increases lung compliance by altering surface properties and airway closure with little change in tissue deformability. Thus, while edema may alter cardiac performance by changing pleural pressure, it has little influence on the pulmonary component of diastolic compliance.

Mechanical cardiopulmonary interdependence.

We studied cardiopulmonary interdependence in ten pentobarbital sodium-anesthetized dogs by 1) measuring the increase of left atrial pressure (Pla) required to hold cardiac output (Q) constant on application of a positive end-expiratory pressure (PEEP), 2) determining the reduction of Pla required to mimic the Q fall observed when PEEP was applied while Pla was held constant, and 3) comparing left ventricular pressure-volume curves measured in freshly dead dogs during ventilation with and without PEEP. The atrial pressure changes can be divided into terms for pleural pressure change, lung deformation, and an undefined residual component and can be used to obtain a compliance opposing ventricular filling. Another compliance was derived from the pressure-volume curves. The latter compliance (6.8 ml/cm H2O) significantly exceeded the former (3.9 ml/cm H2O). The difference may have been caused by ventricular interdependence. The respiratory system compliance opposing ventricular filling was approximately one-twentieth of that predicted from lung and chest wall compliances. Deformation of lungs and chest wall appears to be a significant component of the elastic load imposed on ventricular diastolic filling.