Harjot K. Saini

Prenatal exposure to maternal undernutrition induces adult cardiac dysfunction

An adverse environmental experience of the growing fetus may lead to permanent changes in the structure and function of organs that may predispose the individual to chronic diseases in later life; however, nothing is known about the occurrence and mechanisms of heart failure. We employed a rat model in which pregnant dams were fed diets containing either 180 g (normal) or 90 g (low) casein/kg for 2 weeks before mating and throughout pregnancy. The ejection fraction (EF) of the pups exposed to the low-protein (LP) diet was severely depressed in the first 2 weeks of life and was associated with an increase in cardiomyocyte apoptosis. This early depressed cardiac function was followed by progressive recovery and normalization of the EF of the offspring in the LP group. The left ventricular (LV) internal diameters were increased between 24 h and 84 d (12 weeks) of age in the LP-exposed group. Although between 3 d and 2 weeks of age the LV wall of the heart in the LP group was thinner, a progressive increase in LV wall thickness was seen. At 40 weeks of age, although the EF was normal, a two-fold elevation in LV end-diastolic pressure, reduced cardiac output, decreased maximum rates of contraction and relaxation, and reduced mean arterial pressure were observed. Our findings demonstrate that exposure of the developing fetus to a maternal LP diet programs cardiac dysfunction in the offspring in later life.

Potential role and mechanisms of subcellular remodeling in cardiac dysfunction due to ischemic heart disease.

Several studies have revealed varying degrees of changes in sarcoplasmic reticular and myofibrillar activities, protein content, gene expression and intracellular Ca2+-handling during cardiac dysfunction due to ischemia–reperfusion (I/R); however, relatively little is known about the sarcolemmal and mitochondrial alterations, as well as their mechanisms in the I/R hearts. Because I/R is associated with oxidative stress and intracellular Ca2+-overload, it has been indicated that changes in subcellular activities, protein content and gene expression due to I/R are related to both oxidative stress and Ca2+-overload. Intracellular Ca2+-overload appears to induce changes in subcellular activities, protein contents and gene expression (subcellular remodeling) by activation of proteases and phospholipases, as well as by affecting the genetic apparatus, whereas oxidative stress is considered to cause oxidation of functional groups of different subcellular proteins in addition to modifying the genetic machinery. Ischemic preconditioning, which is known to depress the development of both intracellular Ca2+-overload and oxidative stress due to I/R, was observed to attenuate the I/R-induced subcellular remodeling and improve cardiac performance. It is suggested that a combination therapy with antioxidants and interventions, which reduce the development of intracellular Ca2+-overload, may improve cardiac function by preventing or attenuating the occurrence of subcellular remodeling due to ischemic heart disease. It is proposed that defects in the activities of subcellular organelles may serve as underlying mechanisms for I/R-induced cardiac dysfunction under acute conditions, whereas subcellular remodeling due to alterations in gene expression may explain the impaired cardiac performance under chronic conditions of I/R.

Role of reactive oxygen species in ischemic preconditioning of subcellular organelles in the heart.

Ischemic preconditioning (IPC) is an endogenous adaptive mechanism and is manifested by early and delayed phases of cardioprotection. Brief episodes of ischemia-reperfusion during IPC cause some subtle functional and structural alterations in sarcolemma, mitochondria, sarcoplasmic reticulum, myofibrils, glycocalyx, as well as nucleus, which render these subcellular organelles resistant to subsequent sustained ischemia-reperfusion insult. These changes occur in functional groups of various receptors, cation transporters, cation channels, and contractile and other proteins, and may explain the initial effects of IPC. On the other hand, induction of various transcriptional factors occurs to alter gene expression and structural changes in subcellular organelles and may be responsible for the delayed effects of IPC. Reactive oxygen species (ROS), which are formed during the IPC period, may cause these changes directly and indirectly and act as a trigger of IPC-induced cardioprotection. As ROS may be one of the several triggers proposed for IPC, this discussion is focused on the current knowledge of both ROS-dependent and ROS-independent mechanisms of IPC. Furthermore, some events, which are related to functional preservation of subcellular organelles, are described for a better understanding of the IPC phenomenon.

Pharmacological basis of different targets for the treatment of atherosclerosis

The development of atherosclerotic plaque is a highly regulated and complex process which occurs as a result of structural and functional alterations in endothelial cells, smooth muscle cells (SMCs), monocytes/macrophages, T‐lymphocytes and platelets. The plaque formation in the coronary arteries or rupture of the plaque in the peripheral vasculature in latter stages of atherosclerosis triggers the onset of acute ischemic events involving myocardium. Although lipid lowering with statins has been established as an important therapy for the treatment of atherosclerosis, partially beneficial effects of statins beyond decreasing lipid levels has shifted the focus to develop newer drugs that can affect directly the process of atherosclerosis. Blockade of renin angiotensin system, augmentation of nitric oxide availability, reduction of Ca2+ influx, prevention of oxidative stress as well as attenuation of inflammation, platelet activation and SMC proliferation have been recognized as targets for drug treatment to control the development, progression and management of atherosclerosis. A major challenge for future drug development is to formulate a combination therapy affecting different targets to improve the treatment of atherosclerosis.

Imidapril treatment improves the attenuated inotropic and intracellular calcium responses to ATP in heart failure due to myocardial infarction

1 M Adenosine 5′‐triphosphate (ATP) is known to augment cardiac contractile activity and cause an increase in intracellular Ca2+ concentration ([Ca2+]i) in isolated cardiomyocytes. However, no information regarding the ATP‐mediated signal transduction in the myocardium in congestive heart failure (CHF) is available. 2 CHF due to myocardial infarction (MI) in rats was induced by the occlusion of the left coronary artery for 8 weeks. The positive inotropy due to ATP was depressed in failing hearts. Treatment of 3 weeks infarcted animals with imidapril (1 mg kg−1 day−1) for a period of 5 weeks improved the left ventricle function and decreased the attenuation of inotropic response to ATP. 3 ATP‐induced increase in [Ca2+]i was significantly depressed in cardiomyocytes isolated from the failing heart and this change was partially attenuated by imidapril treatment. However, the binding characteristics of 35S‐labeled adenosine 5′‐(γ‐thio) triphosphate in sarcolemma isolated from the failing heart remained unaltered. 4 ATP‐induced increase in [Ca2+]i was depressed by verapamil and cibacron blue in both control and failing heart cardiomyocytes; however, the ATP response in the failing hearts, unlike the control preparations, was not decreased by ryanodine. This insensitivity to ryanodine was attenuated by imidapril treatment. 5 Treatment of infarcted rats with enalapril and losartan produced effects similar to imidapril. 6 These findings indicate that the positive inotropic response to ATP and ATP‐induced increase in [Ca2+]i in cardiomyocytes are impaired in heart failure. Furthermore, blockade of renin angiotensin system prevented the impairment of the ATP‐mediated inotropic and [Ca2+]i responses in the failing heart.

Changes in β-adrenoceptors in heart failure due to myocardial infarction are attenuated by blockade of renin-angiotensin system

Earlier studies have revealed an improvement of cardiac function in animals with congestive heart failure (CHF) due to myocardial infarction (MI) by treatment with angiotensin converting enzyme (ACE) inhibitors. Since heart failure is also associated with attenuated responses to catecholamines, we examined the effects of imidapril, an ACE inhibitor, on the β-adrenoceptor (β-AR) signal transduction in the failing heart. Heart failure in rats was induced by occluding the coronary artery, and 3 weeks later the animals were treated with 1 mg/(kg·day) (orally) imidapril for 4 weeks. The animals were assessed for their left ventricular function and inotropic responses to isoproterenol. Cardiomyocytes and crude membranes were isolated from the non-ischemic viable left ventricle and examined for the intracellular concentration of Ca2+ [Ca2+]i and β-ARs as well as adenylyl cyclase (AC) activity, respectively. Animals with heart failure exhibited depressions in ventricular function and positive inotropic response to isoproterenol as well as isoproterenol-induced increase in [Ca2+]i in cardiomyocytes; these changes were attenuated by imidapril treatment. Both β1-AR receptor density and isoproterenol-stimulated AC activity were decreased in the failing heart and these alterations were prevented by imidapril treatment. Alterations in cardiac function, positive inotropic effect of isoproterenol, β1-AR density and isoproterenol-stimulated AC activity in the failing heart were also attenuated by treatment with another ACE inhibitor, enalapril and an angiotensin II receptor antagonist, losartan. The results indicate that imidapril not only attenuates cardiac dysfunction but also prevents changes in β-AR signal transduction in CHF due to MI. These beneficial effects are similar to those of enalapril or losartan and thus appear to be due to blockade of the renin–angiotensin system. (Mol Cell Biochem 263: 11–20, 2004)

Mechanisms of low Na(+)-induced increase in intracellular calcium in KCl-depolarized rat cardiomyocytes.

Although low Na+ is known to increase the intracellular Ca2+ concentration ([Ca2+]i) in cardiac muscle, the exact mechanisms of low Na+-induced increases in [Ca2+]i are not completely defined. To gain information in this regard, we examined the effects of low Na+ (35 mM) on freshly isolated cardiomyocytes from rat heart in the absence and presence of different interventions. The [Ca2+]i in cardiomyocytes was measured fluorometrically with Fura-2 AM. Following a 10 min incubation, the low Na+-induced increase in [Ca2+]i was only observed in cardiomyocytes depolarized with 30 mM KCl, but not in quiescent cardiomyocytes. In contrast, low Na+ did not alter the ATP-induced increase in [Ca2+]i in the cardiomyocytes. This increase in [Ca2+]i due to low Na+ and elevated KCl was dependent on the extracellular concentration of Ca2+ (0.25–2.0 mM). The L-type Ca2+-channel blockers, verapamil and diltiazem, at low concentrations (1 μM) depressed the low Na+, KCl-induced increase in [Ca2+]i without significantly affecting the response to low Na+ alone. The low Na+, high KCl-induced increase in [Ca2+]i was attenuated by treatments of cardiomyocytes with high concentrations of both verapamil (5 and 10 μM), and diltiazem (5 and 10 μM) as well as with amiloride (5–20 μM), nickel (1.25–5.0 mM), cyclopiazonic acid (25 and 50 μM) and thapsigargin (10 and 20 μM). On the other hand, this response was augmented by ouabain (1 and 2 mM) and unaltered by 5-(N-methyl-N-isobutyl) amiloride (5 and 10 μM). These data suggest that in addition to the sarcolemmal Na+−Ca2+ exchanger, both sarcolemmal Na+−K+ATPase, as well as the sarcoplasmic reticulum Ca2+-pump play prominent roles in the low Na+-induced increase in [Ca2+]i. (Mol Cell Biochem 263: 151–162, 2004)

Mechanisms of cardiodepression by an Na+-H+ exchange inhibitor methyl-N-isobutyl amiloride (MIA) on the heart: lack of beneficial effects in ischemia-reperfusion injury.

Although Na+-H+ exchange (NHE) inhibitors such as methyl-N-isobutyl amiloride (MIA) are known to depress the cardiac function, the mechanisms of their negative inotropic effect are not completely understood. In this study, isolated rat hearts were perfused with MIA to study its action on cardiac performance, whereas isolated subcellular organelles such as sarcolemma, myofibrils, sarcoplasmic reticulum, and mitochondria were treated with MIA to determine its effect on their function. The effect of MIA on intracellular Ca2+ mobilization was examined in fura-2-AM-loaded cardiomyocytes. MIA was observed to depress cardiac function in a concentration-dependent manner in HCO3- -free buffer. On the other hand, MIA had an initial positive inotropic effect followed by a negative inotropic effect in HCO3-containing buffer. MIA increased the basal concentration of intracellular Ca2+ ([Ca2+]i) and augmented the KCl-mediated increase in [Ca2+]i. MIA did not show any direct effect on myofibrils, sarcolemma, and sarcoplasmic reticulum ATPase activities; however, this agent was found to decrease the intracellular pH, which reduced the myofibrils Ca2+-stimulated ATPase activity. MIA also increased Ca2+ uptake by mitochondria without having any direct effect on sarcoplasmic reticulum Ca2+ uptake. In addition, MIA did not protect the hearts subjected to mild Ca2+ paradox as well as ischemia-reperfusion-mediated injury. These results suggest that the increase in [Ca2+]i in cardiomyocytes may be responsible for the initial positive inotropic effect of MIA, but its negative inotropic action may be due to mitochondrial Ca2+ overloading as well as indirect depression of myofibrillar Ca2+ ATPase activity. Thus the accumulation of [H+]i as well as occurrence of intracellular and mitochondrial Ca2+ overload may explain the lack of beneficial effects of MIA in preventing the ischemia-reperfusion-induced myocardial injury.

Modification of Biochemical and Cellular Processes in the Development of Atherosclerosis by Red Wine

Atherosclerosis is commonly associated with unstable angina and acute myocardial infarction. It occurs as a result of a cascade of events caused by various environmental, dietary, genetic, and inflammatory factors. Different epidemiological studies have suggested that moderate amounts of red wine consumption reduce the risk of complications associated with atherosclerosis. This contention is further supported by a variety of experimental investigations demonstrating that both alcoholic and phenolic components are responsible for the apparent protective effects of red wine. The maintenance of endothelial function, augmentation in the levels of high-density lipoproteins (HDLs), prevention of low-density lipoprotein (LDL) oxidation, attenuation of smooth muscle proliferation and migration, inhibition of platelet aggregation and adhesion, as well as reduction in inflammatory mediators are major mechanisms linked with the protective effects of red wine. Despite these beneficial effects, insufficient information is available to recommend red wine as a therapeutic strategy to prevent atherosclerosis. Particularly, in view of high alcoholic content, excessive consumption of red wine can be seen to produce harmful effects. Therefore, a large-scale clinical trial is needed to determine the exact amount of red wine required for the beneficial effects and to categorize it as a future antiatherosclerotic agent.