Charles F. Pilati

Methamphetamine–gonadal steroid hormonal interactions:: Effects upon acute toxicity and striatal dopamine concentrations

Methamphetamine (MA)-related deaths and nigrostriatal dopaminergic (NSDA) neurotoxicity are greater in males. The exact basis for this gender difference is not known, but data, which show that estrogen (E) can function as a protectant of both the cardiovascular and NSDA systems, suggest an important role for gonadal steroids in modulating toxicity to this psychostimulant. In the present report, we examined the effects of treatment with the gonadal steroid hormones E and testosterone (T) upon MA-induced toxicity within intact and castrated female and male CD-1 mice. Treatment of intact males with E produced a severe acute toxicity to MA, with only 41% (7/17) males surviving at 24-h post-MA. This incidence of mortality was significantly different from that of nonhormonally treated mice receiving an identical regimen of MA [94% survival (16/17)]. None of the other treatment groups showed mortality rates, which differed significantly from the nonhormonally treated mice. Striatal dopamine (DA) concentrations of E-treated female mice (intact or castrated) were significantly greater than that of the nonhormonally treated mice, which failed to differ statistically among each other. In an attempt to understand some of the bases for the mortality rates in E-treated intact males, body temperatures, heart rates and heart catecholamine concentrations were measured from an additional group of intact male mice treated or not treated with E. Heart rates of E-treated intact males were significantly decreased compared with non-E-treated males. No statistically significant differences were obtained for body temperatures or heart catecholamine concentrations. These data demonstrate that MA induces an exacting, acute toxicity, which is specific for E-treated intact male mice and is associated with a reduction in heart rate. In addition, E can function as a neuroprotectant of NSDA system within female, but not male, mice. These data suggest that acute MA toxicity observed with E in intact male mice may result from a change in cardiac function. Accordingly, gonadal steroid hormones can function as critical modulators of both central and peripheral toxicological effects of MA.

Excessive sympathetic nervous system activity decreases myocardial contractility

Abstract The objective of this study was to determine whether myocardial contractility is depressed by intense activation of the sympathetic nervous system. A massive sympathetic discharge was produced by injecting veratrine or sodium citrate into the cisterna magna of anesthetized rabbits (n = 10). Two and one-half hr later, the hearts were isolated and their left ventricular (LV) performance evaluated and compared with the LV performance of hearts isolated from control animals (n = 10). LV performance was evaluated from steady-state peak isovolumic systolic and end-diastolic pressures that were generated at various end-diastolic volumes (LV function curves). The relationship between peak LV systolic pressure (or the average peak developed LV wall stress) and LV end-diastolic volume was rotated downward (P < 0.01) in the hearts removed from rabbits treated with veratrine or citrate. The LV end-diastolic pressure or LV end-diastolic wall stress of these hearts was not different from control at any end-diastolic volume. The diminished ability of the experimental hearts to develop systolic pressure or wall stress suggests that intense sympathetic activation depressed contractility. Severely damaged myofibers, located largely in the subendocardium, were found in these hearts. Furthermore, the depressed contractility was not related to pulmonary edema since only 2 of 10 rabbits developed edema.

β-adrenoceptor stimulation of alveolar fluid clearance is increased in rats with heart failure

The alveolar epithelium plays a critical role in resolving pulmonary edema. We thus hypothesized that its function might be upregulated in rats with heart failure, a condition that severely challenges the lungs ability to maintain fluid balance. Heart failure was induced by left coronary artery ligation. Echocardiographic and cardiovascular hemodynamics confirmed its development at 16 wk postligation. At that time, alveolar fluid clearance was measured by an increase in protein concentration over 1 h of a 5% albumin solution instilled into the lungs. Baseline alveolar fluid clearance was similar in heart failure and age-matched control rats. Terbutaline was added to the instillate to determine whether heart failure rats responded to beta-adrenoceptor stimulation. Alveolar fluid clearance in heart failure rats was increased by 194% after terbutaline stimulation compared with a 153% increase by terbutaline in control rats. To determine the mechanisms responsible for this accelerated alveolar fluid clearance, we measured ion transporter expression (ENaC, Na-K- ATPase, CFTR). No significant upregulation was observed for these ion transporters in the heart failure rats. Lung morphology showed significant alveolar epithelial type II cell hyperplasia in heart failure rats. Thus, alveolar epithelial type II cell hyperplasia is the likely explanation for the increased terbutaline-stimulated alveolar fluid clearance in heart failure rats. These data provide evidence for previously unrecognized mechanisms that can protect against or hasten resolution of alveolar edema in heart failure.

Ouabain-Induced Mechanical Toxicity: Aberrations in Left Ventricular Function, Calcium Concentration, and Ultrastructure

Abstract The isolated rabbit heart was employed to determine if the myocardial mechanical dysfunction (mechanical toxicity) produced by digitalis was accompanied by ultrastructural lesions and an excessive accumulation of calcium by the cardiac cell. Ouabain (1.2 or 2.4 μM) produced a transient increase and then a continuous decrease in the systolic pressure development by the isovolumic left ventricle. The end-diastolic pressure increased progressively during the time that the systolic pressure was diminishing. The ultrastructural abnormalities that were observed in the ouabain-treated hearts included swollen mitochondria, hypercontracted sarcomeres and, occasionally, myofibrillar disruption. The left ventricular cellular calcium concentration was increased (P < 0.05) in these mechanically toxic hearts and this increase correlated significantly (r = 0.65, P < 0.05) with the magnitude of the increase in end-diastolic pressure. Furthermore, a reduction in the perfusate calcium concentration did not attenuate the ouabain-stimulated uptake of calcium. The data suggest that the impaired mechanical performance and the ultrastructural aberrations that may develop in the digitalis-treated heart are related to an excessive uptake of calcium by the left ventricle.

Effects of Cadmium on Contractility and Calcium Concentration in Isolated Heart Muscle

Abstract The isometrically arranged kitten heart papillary muscle was used to study the depressant actions of cadmium ion (Cd2+) on heart muscle function. Although the Cd2+-induced decreases in papillary muscle tension development were dose dependent, peak tension was restored to pretreatment control only after the termination of 1 μM, but not 10 or 100 μM Cd2+ exposure. The Schild plot suggested that Cd2+ and Ca2+ are competitive antagonists but two additional observations indicated that noncompetitive mechanisms are also involved: (a) the contour of the tension twitch from the Cd2+-depressed muscle could not be restored to pre-cadmium control by increasing extracellular Ca2+ concentration, and (b) the contraction pattern after a reduction in external Ca2+ concentration from 2.0 to 0.5 mM differed from that after exposure to 25 μM Cd2+, even though both treatments depressed peak tension development by 80%. When the coronary-perfused rabbit heart was employed, left ventricular Ca concentration was not affected by perfusion with 100 μM Cd2+ even though mechanical function was obliterated by this dose. In conclusion, the mechanisms of action of Cd2+ are complex and do not appear to involve the displacement of Ca from the cardiac cell.

Persistent left ventricular dysfunction after cocaine treatment in rabbits

Abstract The present study was undertaken to determine whether the diminished cardiac performance associated with cocaine administration persists after the drug has been eliminated from the body. Cocaine (5 or 10 mg/kg iv) was administered to conscious (n = 7) or pentobarbital-anesthetized (n = 7) rabbits, respectively. Seven conscious and seven anesthetized control rabbits received the saline vehicle. Two and one-half hours later, the hearts were removed from the animals and perfused under cocaine-free conditions. Left ventricular (LV) contractility was evaluated by plotting steady-state LV systolic and diastolic pressures as a function of LV end-diastolic volume (preload). LV systolic performance was diminished in a dose-related manner in hearts isolated from cocaine-treated rabbits, but was statistically different from control only at the higher cocaine dose (P < 0.05). In a second set of experiments, hearts (n = 6) were isolated, and their LV function was evaluated before, during, and after cocaine exposure. In these experiments, cocaine was added to the perfusate in increments to produce concentrations of 5, 10, and 15 mg/liter. After LV function was evaluated at the highest cocaine dose, cocaine-free perfusion conditions were restored, and LV function was reevaluated. In these experiments, cocaine produced a dose-dependent decrease in LV function that readily reversed when cocaine-free perfusion was reinstated. We conclude that cocaine diminishes LV contractility, and that the diminished cardiac performance may not readily reverse after in vivo exposure. Moreover, the rapid restoration of cardiac performance after exposure to cocaine in vitro suggests that the mechanism operating in vivo involves more than a simple direct action on the myocyte. Catecholamine cardiotoxicity does not appear to be a primary factor.

Effect of intracoronary infusion of histamine or compound 4880 on coronary vascular permeability and myocardial fluid balance

This study was undertaken to determine if coronary vascular permeability (CVP) increases and if myocardial edema develops in the canine heart after local exposure to histamine. Histamine (50 or 15 micrograms/min) or compound 48/80 (0.1-0.2 mg/kg) was infused into the left anterior descending coronary artery (LAD) of open-chest dogs, and changes in CVP were determined by comparing prenodal cardiac lymph flow (Q1) and lymph-to-plasma protein concentration ratio (C1/Cp) before and during histamine or compound 48/80 treatment. CVP increased in most, but not all, experiments with both doses of histamine as indicated by simultaneous increases in both Q1 and C1/Cp. The injection of compound 48/80 into the LAD of four dogs caused unequivocal increases in CVP in only one experiment. Compared with the effect of histamine on the forelimb, the average increases in Q1 and C1/Cp were not large with either histamine or compound 48/80, which suggests that the increases in CVP were relatively small. Moreover, edema did not develop. These results indicate that the coronary microvasculature of the intact dog heart is relatively insensitive to increases in permeability produced by histamine. Furthermore, the release of histamine from myocardial mast cells would not be expected to play a major role in the myocardial edema that develops under various pathological conditions.