Intra-uterine growth retardation results in increased cardiac arrhythmias and raised diastolic blood pressure in adult rats.

OBJECTIVES: Epidemiological evidence in humans suggests that intrauterine growth retardation is associated with an increased risk of hypertension and coronary heart disease in later life. To begin to understand the mechanisms involved, we developed and exploited a rat model of intrauterine growth retardation to assess predisposition to arrhythmias and resting blood pressure levels at defined ages from 4 to 18 months. METHODS: Isolated working heart experiments were carried out on rats that had been subjected to intrauterine growth retardation by prenatal protein deprivation and age-matched male Wistar controls to measure susceptibility to wall stress-induced arrhythmias. In addition, resting systolic and diastolic blood pressures were measured in conscious rats via an indwelling arterial catheter. RESULTS: Hearts from intrauterine growth retarded animals showed significantly more ventricular premature beats and more episodes of ventricular tachycardia at all ages examined (4, 9 and 18 months), and at 4 and 18 months, a reduction in coronary blood flow. Diastolic pressure was significantly raised by intrauterine growth retardation in both groups examined (4 and 9 months). CONCLUSIONS: Protein malnutrition during the intrauterine period results in profound intrauterine growth retardation that is associated with a raised diastolic blood pressure and an increased predisposition to cardiac arrhythmias in later life. These results are consistent with epidemiological observations made in human populations, and as similar pathophysiological changes may operate in both situations, intrauterine protein deprivation may be a useful model to help define some of the mechanisms involved.

Changes in contractile protein gene expression with ageing and with captopril-induced regression of hypertrophy in the spontaneously hypertensive rats.

BACKGROUND: Left ventricular hypertrophy (LVH) is present in young spontaneously hypertensive rats (SHR) compared with normotensive Wistar-Kyoto (WKY) rats and treatment of SHR with captopril leads to regression of LVH. Hypertrophy produces changes in gene expression for myofibrillar proteins with increased ratios of skeletal to cardiac actin and beta to alpha-myosin heavy chain (MHC). OBJECTIVES: The objective of this study was to follow changes in transcript prevalence for these four proteins during ageing and with captopril treatment in SHR and WKY rats. METHODS: Untreated SHR and WKY rats were studied at 100, 156, 350 and 450 days. Groups at 100 and 350 days were divided into a treatment group (given captopril) and untreated controls. Transcripts were measured using in situ hybridization. RESULTS: Both cardiac and skeletal actin were increased in untreated SHR compared to WKY rats (P<0.01 and P<0.05, respectively). alpha-MHC was increased (P<0.01) whilst beta-MHC was normal in 100-day-old SHR (an age when LVH was present) compared with WKY rats. With ageing, alpha-MHC declined and beta-MHC increased giving the increased ratio of beta to alpha-MHC transcripts reported by other investigators. Treatment of SHR led to a significant decline in skeletal actin transcripts (P< 0.01) and reversed the rise in beta-MHC expression that occurred with ageing (P< 0.01). CONCLUSIONS: LVH in SHR is associated with increased skeletal and cardiac actin transcripts. Despite unequivocal LVH in SHR at 100 days of age, alpha rather than beta-MHC transcripts were increased. Only with ageing did the classically reported increased ratio of beta to alpha-MHC transcripts become apparent Captopril treatment reduced skeletal actin transcripts and reversed the increase in beta-MHC that occurred with ageing.

Experimental studies on myocardial stretch and ventricular arrhythmia in hypertrophied and non-hypertrophied hearts.

Hypertension affects about 5% of western populations and in the majority of cases it is of unknown aetiology. It exposes the heart to greater levels of myocardial stretch as a result of increased systolic pressure and peripheral resistance. Under certain circumstances myocardial stretch may trigger arrhythmias but the mechanisms and clinical importance of this phenomenon are unclear. This article outlines the risks of sudden cardiac death conferred by hypertension and left ventricular hypertrophy, presents the results of experiments using an animal model of myocardial stretch and discusses some possible mechanisms underlying stretch-induced arrhythmias which may be important in hypertensive patients.

Effect of streptomycin on wall-stress-induced arrhythmias in the working rat heart.

OBJECTIVE: To assess whether streptomycin, an inhibitor of mechano-sensitive cation channels, has an effect on arrhythmias-induced by an increase of ventricular wall stress in the rat heart. METHODS: The isolated working rat heart preparation was used. Arrhythmias were induced by increasing the afterload (i.e., aortic pressure) against which the left ventricle (LV) pumped for 20 s. This led to an increase of LV pressure, stretch of the LV and an increase in LV wall stress. The number of ventricular premature beats induced by each afterload step was compared in the absence and presence of streptomycin, a compound known to block mechano-sensitive cation channels in the heart. RESULTS: Perfusion with 200 microM streptomycin caused a significant reduction in wall-stress-induced arrhythmias. The effect of streptomycin on arrhythmias reached steady-state within 10 min of application. In the presence of streptomycin, arrhythmias elicited by a 40 mmHg afterload increase were reduced to 38% of control. Arrhythmias induced by an 80 mmHg afterload increase were reduced to 61% of control. Complex arrhythmias (ventricular tachycardia) induced by an afterload increase were also reduced in the presence of 200 microM streptomycin. There was no change in inotropic state with streptomycin, as assessed either by cardiac output or by maximum developed LV pressure. Streptomycin 50 microM (a typical therapeutic plasma concentration in patients) had no effect on wall-stress-induced arrhythmias. CONCLUSIONS: The results were inconsistent with streptomycin acting by modulating inositol phosphate production, or altering the level of intracellular calcium or inotropic state. The anti-arrhythmic effect of streptomycin appears more consistent with inhibition of mechano-sensitive cation channels, suggesting that these ion channels might be involved in causing wall-stress-induced arrhythmias.

Role of intracellular sodium overload in the genesis of cardiac arrhythmias.

A number of clinical cardiac disorders may be associated with a rise of the intracellular Na concentration (Na(i)) in heart muscle. A clear example is digitalis toxicity, in which excessive inhibition of the Na/K pump causes the Na(i) concentration to become raised above the normal level. Especially in digitalis toxicity, but also in many other situations, the rise of Na(i) may be an important (or contributory) cause of increased cardiac arrhythmias. In this review, we consider the mechanisms by which a raised Na(i) may cause cardiac arrhythmias. First, we describe the factors that regulate Na(i), and we demonstrate that the equilibrium level of Na(i) is determined by a balance between Na entry into the cell, and Na extrusion from the cell. A number of mechanisms are responsible for Na entry into the cell, whereas the Na/K pump appears to be the main mechanism for Na extrusion. We then consider the processes by which an increased level of Nai might contribute to cardiac arrhythmias. A rise of Na(i) is well known to result in an increase of intracellular Ca, via the important and influential Na/Ca exchange mechanism in the cell membrane of cardiac muscle cells. A rise of intracellular Ca modulates the activity of a number of sarcolemmal ion channels and affects release of intracellular Ca from the sarcoplasmic reticulum, all of which might be involved in causing arrhythmia. It is possible that the increase in contractile force that results from the rise of intracellular Ca may initiate or exacerbate arrhythmia, since this will increase wall stress and energy demands in the ventricle, and an increase in wall stress may be arrhythmogenic. In addition, the rise of Na(i) is anticipated to modulate directly a number of ion channels and to affect the regulation of intracellular pH, which also may be involved in causing arrhythmia. We also present experiments in this review, carried out on the working rat heart preparation, which suggest that a rise of Na(i) causes an increase of wall stress-induced arrhythmia in this model. In addition, we have investigated the effect on wall stress-induced arrhythmia of maneuvers that might be anticipated to change intracellular Ca, and this has allowed identification of some of the factors involved in causing arrhythmia in the working rat heart.

Wall stress-induced arrhythmias in the working rat heart as left ventricular hypertrophy regresses during captopril treatment.

OBJECTIVES: (1) To determine whether regression of left ventricular hypertrophy (LVH) leads to a reduction in wall stress induced arrhythmia. (2) To determine the relationship between the time course of LVH regression and changes in arrhythmias in the spontaneously hypertensive rat heart. METHODS: 67 male spontaneously hypertensive rats (SHR) and 67 normotensive Wistar Kyoto rats (WKY) rats were studied at 100 days of age. 39 of each were treated with the ACE inhibitor captopril 2 mg/ml in drinking water, and the remaining 28 were controls. At 0, 2, 4, 8 and 16 weeks hearts were removed and perfused in the working heart mode. Control afterload was 80 mm Hg and perfusate K+ was 2.4 mM. Step increases in afterload (20, 40 and 80 mm Hg rises; 20 s duration; 2 min between each) were applied in random order to increase ventricular wall stress and induce arrhythmias. RESULTS: Total number of ventricular premature beats (VPBs) elicited by each afterload step were counted. The ratio of left ventricular weight to body weight in the SHR (an index of LVH) showed a rapid and marked decline with captopril treatment (2.65 +/- s.e.m. 0.07 mg/g after 2 weeks treatment compared to 3.38 +/- 0.08 before treatment; P < 0.01), indicating that captopril produced rapid regression of LVH. In contrast, the number of wall stress-induced arrhythmias in SHR did not show a significant decline over the 16 week treatment period. However, when the effect of regression of LVH on wall thickness was taken into account, and compensation was made for differences in wall stress applied, there did appear to be a slow reduction in arrhythmias in SHR. This decline in VPBs was significant after 16 weeks treatment for 40 and 80 mm Hg rises in afterload (P < 0.05). CONCLUSIONS: Treatment with captopril produced a rapid regression of LVH in the SHR. In contrast, arrhythmias declined more slowly over the 16 week period. There did not appear to be a direct relationship between the degree of regression of LVH and wall stress-induced arrhythmias in this model.

"Voltage-activated Ca release" in rabbit, rat and guinea-pig cardiac myocytes, and modulation by internal cAMP.

It is widely believed that Ca release from the sarcoplasmic reticulum (SR) in heart muscle is due to "Ca-induced Ca-release" (CICR), triggered by transmembrane Ca entry. However, in intact guinea-pig cells or cells dialysed with cAMP there may be an additional mechanism - SR release may be activated directly by membrane depolarisation without Ca entry. The first objective of the present study was to investigate whether this "voltage-activated Ca release" (VACR) mechanism is present across species such as rabbit, rat and guinea-pig. The second objective was to characterise the dependence of a VACR mechanism on internal [cAMP]. Membrane current was measured with the whole-cell patch-clamp technique, intracellular [Ca] was monitored with Fura-2 (or a combination of Fluo-3/SNARF-1). Rapid changes of superfusate (within 100 ms) were made using a system which maintained cell temperature at 37 degrees C. We used a train of conditioning pulses to ensure a standard SR load before each test pulse. In rabbit myocytes dialysed with 100 microM cAMP, 89.6 +/- 7.0% of the control intracellular Ca (Cai) transient was still elicited by depolarisation during a switch to 5 mM Ni, which blocked pathways for Ca entry. This suggested that rabbit myocytes possess a VACR mechanism. The percentage of control Cai transient elicited by depolarisation in the presence of 5 mM Ni (i.e. magnitude of VACR) increased in a graded fashion with the pipette [cAMP] between zero and 100 microM. In rat myocytes dialysed with 50 microM cAMP, 64.4 +/- 6.2% of SR release was activated by depolarisation in the presence of 5 mM Ni, suggesting the presence of a VACR mechanism. The extent to which VACR triggered SR release increased with the pipette [cAMP] between zero and 50 microM. In guinea-pig myocytes dialysed with 100 microM cAMP, 74.6 +/- 3.6% of the control Cai transient was elicited by depolarisation in the presence of 5 mM Ni. The degree to which VACR triggered SR release was also graded with the pipette [cAMP] between zero and 100 microM. It therefore appears that each of the three species might possess a VACR mechanism which can be modulated by the internal [cAMP]. This may reflect an effect of cAMP to phosphorylate key proteins involved in excitation-contraction coupling. Under normal physiological conditions with a basal [cAMP] between 2 and 20 microM, VACR may play a role in triggering SR release. The role of VACR may increase under conditions which increase internal [cAMP].