Ken S. Dhalla

Beneficial Effects of Verapamil in Diabetic Cardiomyopathy

It has been suggested that the occurrence of an intracellular Ca2+ overload may result in the development of diabetic cardiomyopathy, which is associated with depletion of high-energy phosphate stores and a derangement of ultrastructure and cardiac dysfunction. Accordingly, the effects of verapamil, a Ca2+ antagonist, on cardiac function, ultrastructure, and high-energy phosphate stores in the myocardium were evaluated in rats made diabetic by an intravenous injection of streptozocin (65 mg/kg). Four weeks after the induction of diabetes, the animals were treated with three doses (2, 4, or 8 mg kg−1 day−1) of verapamil for 4 wk until they were used for the measurement of different parameters. Untreated diabetic animals had slower heart rates, depressed rate of contraction and rate of relaxation, lower peak left ventricular systolic pressure, and elevated left ventricular diastolic pressure. All of these changes were significantly improved in diabetic rats receiving verapamil treatment. The beneficial effects of verapamil were more evident with higher doses (8 mg · kg−1 · day−1) than with the lower doses (2 mg · kg−1 · day−1). The diabetic animals also showed alterations in myocardial high-energy phosphate stores and exhibited evidence of ultrastructural damage; these abnormalities were improved by verapamil treatment without affecting their hyperglycemic status. Our results demonstrate that verapamil is capable of preventing diabetes-induced myocardial changes and support the involvement of Ca2+ in the cardiac pathology during diabetes.

Mechanisms of alterations in cardiac membrane Ca2+ transport due to excess catecholamines

SummaryThe occurrence of excessive catecholamine release is often associated with stress due to the lifestyle of Western societies. Contrary to the general thinking that excess catecholamines produce cardiotoxicity mainly via binding to adrenoceptors, there is increasing evidence that catecholamine-induced deleterious actions may also occur through oxidative mechanisms. In this overview it is shown that a high dose of isoproterenol induces a biphasic change in cardiac Ca2+ transport in the sarcolemma and in sarcoplasmic reticulum. Both sarcolemmal and sarcoplasmic reticular Ca2+-transport activities are initially increased to maintain Ca2+ homeostasis and then are impaired, which may be associated with the occurrence of intracellular Ca2+ overload. On the other hand, mitochondrial Ca2+-transport activities exhibited a delayed increase. Pretreatment with vitamin E partially prevented the deleterious changes in cardiac membranes as well as the depressed energetic status of the heart muscle cell. It is concluded that excess catecholamines affect Ca2+-transport mechanisms primarily via oxidation reactions involving free radical-mediated damage. Thus drug approaches that reduce circulating catecholamines and/or prevent their oxidation should prove beneficial. A combination therapy involving inhibitors of catecholamine release, blockers of adrenoceptors, and antioxidants may be indicated for stress-induced heart disease.

Mechanisms of cardiac cell damage due to catecholamines: significance of drugs regulating central sympathetic outflow.

A chronically increased rate of catecholamine release has various deleterious actions. Isoproterenol injections (80 mg/kg body weight) resulted in depressed Ca2+ transport in the sarcolemma (ATP-dependent Ca2+ uptake, Na(+)-dependent Ca2+ uptake) and sarcoplasmic reticulum (Ca2+ uptake) of rat heart. The formation of malondialdehyde owing to lipid peroxidation was increased. Pretreatment with vitamin E (10-25 mg/kg/day) strongly inhibited the membrane damage. The toxic effects of catecholamines arise most probably from their oxidation, and it is therefore important either to reduce the central sympathetic outflow or to prevent the oxidation. An inappropriately high sympathetic outflow is a typical feature of Western affluent societies, and is linked to psychosocial stress and hypercaloric nutrition. However, established pharmacologic interventions to reduce sympathetic outflow have proven not practicable because of marked side effects. Using radiotelemetry for monitoring cardiovascular parameters of spontaneously hypertensive rats treated with clonidine or moxonidine, we showed that clonidine, unlike moxonidine, resulted in rebound hypertension after drug withdrawal. Because the rebound blood pressure and the typical side effects of clonidine associated with low patient compliance are mainly mediated by alpha-adrenoceptors, it can be inferred that the I1-imidazoline agonist moxonidine does not exhibit the side effects commonly seen with clonidine and therefore represents a promising approach for reducing an inappropriately high central sympathetic outflow.

Measurement of adrenolutin as an oxidation product of catecholamines in plasma

Using the reverse phase high-performance liquid chromatography (HPLC) with mobile phases composed of simple acids, we have developed an assay technique for the measurement of adrenolutin, one of the oxidation products of catecholamines, in rat plasma. Ion-pairing chromatography permits the separation and quantitation of plasma adrenolutin (μM) in a linear manner. Sample preparation involved the precipitation of plasma proteins with perchloric acid and it is easier to handle a large number of samples at a time. However, we were unable to demonstrate the presence of adrenochrome, another oxidation product of catecholamines, in plasma since adrenochrome was rapidly destroyed in acid as well as in blood and was quickly changed, into adrenolutin. Adrenolutin peak in HPLC was confirmed by 1) the retention time; 2) co-injection of adrenolutin and; 3) the appearance of 3H-adrenolutin after injection of 3H-norepinephrine. Administration of different catecholamines as well as adrenochrome and adrenolutin in rats also increased the level of adrenolutin in plasma. Adrenolutin was found to be present in plasma in other species including dog, rabbit and pig. High level of adrenolutin, which may represent total concentration of aminolutin in plasma, suggests the presence of an efficient mechanism for the oxidation of catecholamines under in vivo conditions.

Differential changes in sympathetic activity in left and right ventricles in congestive heart failure after myocardial infarction

Although congestive heart failure subsequent to myocardial infarction is known to be associated with increased sympathetic activity, very little information regarding changes in the sympathetic nerves in the left and right ventricles at various stages after infarction is available. Male Sprague-Dawley rats were subjected to coronary artery ligation and studied 4 and 8 weeks later; these animals had mild and moderate stages of congestive heart failure. A sham group, without coronary ligation, was used as control. Four weeks after myocardial infarction, plasma and ventricular (left and right) epinephrine (EPI), unlike norepinephrine (NE), were markedly increased. Whereas plasma catecholamine (EPI and NE) levels were increased 8 weeks after infarction, NE concentration in the left ventricle was unchanged but EPI concentration was increased in comparison with sham control. The right ventricle showed an increased level of both NE and EPI 8 weeks after infarction. Measurement of the rate of change in the specific activity of NE (NE turnover) in the left and right ventricles 8 weeks after infarction revealed an increase in NE turnover in the left ventricle, without any changes in the right ventricle. The concentration of EPI, unlike NE, was increased in the kidney, spleen, and brain 8 weeks after coronary occlusion. These results are interpreted to mean that congestive heart failure caused by myocardial infarction is associated with differential changes in the status of sympathetic nerves in the left and right ventricles; sympathetic activity is increased only in the left ventricle, whereas the right ventricle may play an adaptive role by increasing catecholamine stores during the development of heart failure.

Effects of long-term dietary restriction on cardiovascular function and plasma catecholamines in the rat

SummaryTo examine the relationship between heart function and plasma catecholamines upon food restriction, normal adult rats were fed 12 g or 6 g food/day for 14 days and 12 g food/day for 28 days. Food-restricted rats exhibited bradycardia, hypotension, and decreased rates of cardiac contraction (+dP/dt) as well as relaxation (-dP/dt) at 14 (12 or 6 g food/day) and 28 (12 g food/day) days. Plasma norepinephrine and epinephrine levels were significantly elevated in the 6 g food/day group at 14 days, whereas in the 12 g food/day group, plasma norepinephrine was elevated at 14 days but was significantly decreased at 28 days. Heart norepinephrine and epinephrine concentrations were elevated at both 14 and 28 days of food restriction in the 12 g food/day group as well as at 14 days in the 6 g food/day group. Thus, dietary restriction appears to result in depressed indices of heart function, while the circulating levels of catecholamines were elevated at early states.

Plasma neuropeptide Y in anxiety disorders: findings in panic disorder and social phobia.

The demonstration in preclinical studies that centrally administered neuropeptide Y (NPY) has anxiolytic effects had led to speculation that NPY may play a role in human anxiety disorders. We therefore decided to study plasma NPY levels in 22 patients with DSM-III-R anxiety disorders (11 with panic disorder and 11 with social phobia, generalized type) and 12 never psychiatrically ill comparison subjects. Under resting conditions, plasma NPY levels did not differ among the three diagnostic groups. Following hand immersion in ice water, plasma NE levels--but not NPY levels--increased immediately, but there were no significant differential diagnostic effects. These results are convergent with prior reports of normal sympathetic nerve activity in patients with anxiety disorders.

Status of Post Adrenergic Receptor Mechanisms in Cardiac Hypertrophy and Heart Failure

It is now well known that heart function is regulated by the sympathetic nervous system, in which norepinephrine released from the nerve endings activates J3-adrenergic receptors in the myocardium and increases the development of contractile force [1,2]. The activation of the contractile apparatus by catecholamines occurs through the formation of cyclic AMP and a subsequent increase in the intracellular concentration of Ca2+. At the biochemical level, this signal transduction pathway, in which β-adrenoceptors are coupled to adenylyl cyclase through guanine nucleotide binding proteins (G-proteins), is considered to influence cardiac contraction and relaxation processes by phosphorylation of various membrane and contractile proteins. A schematic representation of events involved in the β-adrenergic receptor mechanisms, leading to increased heart function, is shown in Figure 1.