Alterations in excitation–contraction coupling and elevated intracellular sodium (Nai) are hallmarks of pathological cardiac remodelling that underline contractile dysfunction. In addition, changes in cardiac metabolism are observed in cardiac hypertrophy and heart failure (HF) that lead to a mismatch in ATP supply and demand, contributing to poor prognosis. A link between Nai and altered metabolism has been proposed but is not well understood. Many mitochondrial enzymes are stimulated by mitochondrial calcium (Camito) during contraction, thereby sustaining production of reducing equivalents to maintain ATP supply. This stimulation is thought to be perturbed when cytosolic Nai is high due to increased Camito efflux, potentially compromising ATPmito production and leading to metabolic dysregulation. Increased Nai has been previously shown to affect Camito; however, whether Nai elevation plays a causative role in energetic mismatching in the hypertrophied and failing heart remains unknown. In this review, we discuss the relationship between elevated Nai, NaK ATPase dysregulation and the metabolic phenotype in the contexts of pathological hypertrophy and HF and their link to metabolic flexibility, capacity (reserve) and efficiency that are governed by intracellular ion homeostasis. The development of non-invasive analytical techniques using nuclear magnetic resonance able to probe metabolism in situ in the functioning heart will enable a better understanding of the underlying mechanisms of Nai overload in cardiac pathophysiology. They will lead to novel insights that help to explain the metabolic contribution towards these diseases, the incomplete rescue observed with current therapies and a rationale for future energy-targeted therapies.

Cardiovascular disease is the leading cause of mortality worldwide with its incidence projected to rise significantly in the immediate future. There is a clear need for improved understanding of underlying cellular mechanisms which can aid the development of more effective treatments as well as novel techniques for early diagnosis. There is a convincing evidence that myocardial intracellular Na (Nai) overload along with metabolic derangement are two important and interconnected pathophysiological features of hypertrophy and heart failure (HF). Na ion homeostasis is regulated by many transporters and membrane pumps [1]. Na/K ATPase (NKA) and its key regulatory protein phospholemman (PLM) play a crucial role in cardiomyocyte transmembrane ion transport and contractility, such that transgenic PLM3SA mice, in which PLM is rendered unphosphorylatable, have chronically elevated Nai and an increased susceptibility to hypertrophy-induced dysfunction [2]. In addition, due to the high ATP demand required for pump activity and its sarcolemmal localization, there is evidence for an association between NKA and metabolism with its ATP supply thought to be supplied preferentially by glycolysis [35]. More recent studies have suggested that cytosolic Na regulation also plays an important role in linking mitochondrial Ca-dependent ATP production to mechanical activity and ATP demand due to contractile work [6,8]. Nevertheless, the extent that these metabolic alterations (mismatch in ATP supply–demand) reflect chronic cellular remodelling or arise as a consequence of Nai elevation is not well understood.

In most larger mammalian hearts, with a long action potential, Nai is maintained at ∼4–8 mM [9,10]. In murinae (rats and mice), intracellular Na is significantly elevated (10–20 mM) and this elevated Na is associated with many other adaptations in excitation–contraction (EC) coupling, including a short action potential, a larger recirculating Ca fraction, a dependence on SR Ca release, reduced NCX (sarcolemmal sodium–calcium exchanger) activity, rest potentiation and a negative force–frequency staircase [6].

The cell exploits the energy in the transmembrane Na gradient to drive a plethora of Na-dependent membrane transporters moving ions, substrates, amino acids etc. either into (co-transporters/symports) or out of (exchangers/antiports) the cell (Figure 1). The importance of this trans-sarcolemmal inward Na gradient means that its dissipation in various pathologies such as ischaemia/reperfusion [11], hypertrophy or HF [12,13] is highly detrimental. While some of the Na transport processes are electro-neutral, some are electrogenic and hence both respond to, and contribute to, the membrane potential. Most notably, voltage-gated Na channels are crucially important in generating the upstroke of the cardiac action potential. While there are a large number of Na influx pathways, there is only a single quantitatively significant Na efflux pathway responsible for maintaining the transmembrane Na gradient — the Na/K ATPase or Na/K pump (NKA) [14].

Major Nai influx and efflux pathways and metabolic pathways involved in ATP supply.

Figure 1.
Major Nai influx and efflux pathways and metabolic pathways involved in ATP supply.

The delivery of metabolic substrates, their selection and uptake are followed by OXPHOS. It involves electron shuttling from cytosolic to mitochondrial reducing equivalents, transfer of energy by electrons from reducing equivalents to ETC complexes and generation of electrochemical proton (H+) gradient within the mitochondrial intermembrane space (respiratory complexes I, II, II, III, IV). The release of H+ gradient is coupled to the synthesis of ATP from ADP + Pi by F0,F1-ATPase (complex V), contributing >95% of ATP synthesis under aerobic conditions. The final stage of myocardial ATP supply (phosphotransfer) involves delivery of ATP from mitochondria to sites of use. This involves ADP–ATP exchange across the inner mitochondrial membrane by the adenine nucleotide transporter (ANT) and propagation of local ATP/ADP disequilibria primarily by the creatine kinase (CK). Abbreviations: TAG, triacylglycerol; PCr, phosphocreatine; ANT, adenine nucleotide transporter; GLUT, glucose transporter; CD36, fatty acid transporter; PPP, pentose phosphate pathway; LDH, lactate dehydrogenase; PDH, pyruvate dehydrogenase; CPT, carnitine palmitoyltransferase; CACT, carnitine–acylcarnitine translocase; MCU, mitochondrial calcium uniporter; α-KDH, α-ketoglutarate dehydrogenase; IDH, isocitrate dehydrogenase; mitoCK, mitochondrial creatine kinase; IMM, inner mitochondrial membrane; OMM, outer mitochondrial membrane; Q, quinone pool; c, cytochrome c; MPC, mitochondrial pyruvate carrier; e, electrons; CGP, mitochondrial Na–Ca exchanger inhibitor CGP-37157. *Mitochondrial calcium-sensitive dehydrogenases (pyruvate dehydrogenase, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase).

Figure 1.
Major Nai influx and efflux pathways and metabolic pathways involved in ATP supply.

The delivery of metabolic substrates, their selection and uptake are followed by OXPHOS. It involves electron shuttling from cytosolic to mitochondrial reducing equivalents, transfer of energy by electrons from reducing equivalents to ETC complexes and generation of electrochemical proton (H+) gradient within the mitochondrial intermembrane space (respiratory complexes I, II, II, III, IV). The release of H+ gradient is coupled to the synthesis of ATP from ADP + Pi by F0,F1-ATPase (complex V), contributing >95% of ATP synthesis under aerobic conditions. The final stage of myocardial ATP supply (phosphotransfer) involves delivery of ATP from mitochondria to sites of use. This involves ADP–ATP exchange across the inner mitochondrial membrane by the adenine nucleotide transporter (ANT) and propagation of local ATP/ADP disequilibria primarily by the creatine kinase (CK). Abbreviations: TAG, triacylglycerol; PCr, phosphocreatine; ANT, adenine nucleotide transporter; GLUT, glucose transporter; CD36, fatty acid transporter; PPP, pentose phosphate pathway; LDH, lactate dehydrogenase; PDH, pyruvate dehydrogenase; CPT, carnitine palmitoyltransferase; CACT, carnitine–acylcarnitine translocase; MCU, mitochondrial calcium uniporter; α-KDH, α-ketoglutarate dehydrogenase; IDH, isocitrate dehydrogenase; mitoCK, mitochondrial creatine kinase; IMM, inner mitochondrial membrane; OMM, outer mitochondrial membrane; Q, quinone pool; c, cytochrome c; MPC, mitochondrial pyruvate carrier; e, electrons; CGP, mitochondrial Na–Ca exchanger inhibitor CGP-37157. *Mitochondrial calcium-sensitive dehydrogenases (pyruvate dehydrogenase, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase).

Close modal

The activity of the NKA is regulated by FXYD1, or PLM, the principal sarcolemmal target of protein kinases A and C (Figure 2) [15]. As such, PLM is required for the dynamic control of Nai during increases in heart rate or during disease and plays a vital role in Na regulation during ‘fight or flight’ [14]. Under physiological conditions, NKA is the only quantitatively significant efflux pathway of Na out of the myocyte (NCX and Na/HCO3/Cl symporter, in principle, can reverse and efflux Na) [16] (Figure 1).

Schematic depiction of a structure–function relationship (regulation) between PLM and Na pump.

Figure 2.
Schematic depiction of a structure–function relationship (regulation) between PLM and Na pump.

The cytoplasmic tail of unphosphorylated PLM interacts closely with the membrane and α-subunit of Na pump, whereas phosphorylation alters the association between the pump and PLM by moving the cytosolic arm away from the pump, but not by promoting their dissociation. Phosphorylation or ablation of PLM relieves inhibition of the Na pump by increasing its Vmax and apparent Na affinity. Under stress, phosphorylation of PLM allows the heart to reduce its Na and Ca load and prevents lethal arrhythmias. Adapted from Pavlovic et al. [15].

Figure 2.
Schematic depiction of a structure–function relationship (regulation) between PLM and Na pump.

The cytoplasmic tail of unphosphorylated PLM interacts closely with the membrane and α-subunit of Na pump, whereas phosphorylation alters the association between the pump and PLM by moving the cytosolic arm away from the pump, but not by promoting their dissociation. Phosphorylation or ablation of PLM relieves inhibition of the Na pump by increasing its Vmax and apparent Na affinity. Under stress, phosphorylation of PLM allows the heart to reduce its Na and Ca load and prevents lethal arrhythmias. Adapted from Pavlovic et al. [15].

Close modal

A hallmark of cardiac hypertrophy and failure is an elevation of Nai. There is an abundant literature on this phenomenon, although absolute values of measured Nai are often dissimilar, probably owing to methodological differences as summarized in Table 1. Elevation in Nai may contribute to the negative force–frequency relationship, slowed relaxation and arrhythmias [17]. While a component of the elevation of Nai may reflect an increase in Na influx [10], there is a large body of evidence showing that Na/K pump function may also be compromised [2,12,13,18]. Specifically, in cardiac hypertrophy, many studies have shown that NKA pump function, and/or expression, is reduced [2,12,13,18,19]. Cardiac Nai can also be elevated by several other factors such as hypothermia [16] or by increased intracellular pH via enhanced NHE (sodium–proton exchanger) activity, for example, following ischaemia–reperfusion injury [20].

Table 1
Summary of studies quantifying bulk cytosolic [Na]i in the myocyte under both physiological and pathophysiological conditions across various mammalian species
Species[Na]i (mM)Method usedReference
Human 8.0 SBFI (sodium-binding benzofuran isophthalate)-loaded muscle strips paced at 0.25 Hz [17
Human LVH 14.2 Na-selective microelectrodes; muscle strips at rest [67
Human failing 12.1 SBFI-loaded muscle strips paced at 0.25 Hz [17
Human MVD 11.8 Na-selective microelectrodes; muscle strips at rest [67
Sheep 5–6.4 Na-selective microelectrodes; Purkinje fibres at 1 Hz and at rest [68,69
5.8–7.9 Na-selective microelectrodes; muscle strips at rest [70
Dog 8.9–10.4 Na-selective microelectrodes; Purkinje fibres at rest and 1 Hz [71
Guinea pig 4.7–8.0 Na-selective microelectrodes; muscle strips at rest [67,70,72,73,74
6.4 23Na NMR; isolated perfused heart [75
5.1–5.2 SBFI-loaded myocytes at rest [9,76
Guinea pig LVH 12.1 Na-selective microelectrodes; muscle strips at rest [67
12.8 23Na NMR; isolated perfused heart [75
Guinea pig failing 16.8 SBFI-loaded myocytes at rest [77
Ferret 7.8 Na-selective microelectrodes; muscle strips at rest [78
Ferret RVH 8.0 Na-selective microelectrodes; muscle strips at rest [78
Rabbit 7.2 Na-selective microelectrodes; muscle strips at 0.5 Hz [6,10
3.8–4.5 SBFI-loaded myocytes at rest [10,79
Rat 12.7 Na-selective microelectrodes; muscle strips at 0.5 Hz [6
8.5–30 Na-selective microelectrodes; myocytes at rest [80
5.1–21 SBFI-loaded myocytes at rest [9,10,79,81
17.5 23Na NMR; isolated perfused arrested hearts [82
Mouse 11.6 23Na NMR; isolated perfused heart [51
Mouse 14 SBFI-loaded myocytes at rest [2
Mouse LVH 23 SBFI-loaded myocytes at rest [2
Species[Na]i (mM)Method usedReference
Human 8.0 SBFI (sodium-binding benzofuran isophthalate)-loaded muscle strips paced at 0.25 Hz [17
Human LVH 14.2 Na-selective microelectrodes; muscle strips at rest [67
Human failing 12.1 SBFI-loaded muscle strips paced at 0.25 Hz [17
Human MVD 11.8 Na-selective microelectrodes; muscle strips at rest [67
Sheep 5–6.4 Na-selective microelectrodes; Purkinje fibres at 1 Hz and at rest [68,69
5.8–7.9 Na-selective microelectrodes; muscle strips at rest [70
Dog 8.9–10.4 Na-selective microelectrodes; Purkinje fibres at rest and 1 Hz [71
Guinea pig 4.7–8.0 Na-selective microelectrodes; muscle strips at rest [67,70,72,73,74
6.4 23Na NMR; isolated perfused heart [75
5.1–5.2 SBFI-loaded myocytes at rest [9,76
Guinea pig LVH 12.1 Na-selective microelectrodes; muscle strips at rest [67
12.8 23Na NMR; isolated perfused heart [75
Guinea pig failing 16.8 SBFI-loaded myocytes at rest [77
Ferret 7.8 Na-selective microelectrodes; muscle strips at rest [78
Ferret RVH 8.0 Na-selective microelectrodes; muscle strips at rest [78
Rabbit 7.2 Na-selective microelectrodes; muscle strips at 0.5 Hz [6,10
3.8–4.5 SBFI-loaded myocytes at rest [10,79
Rat 12.7 Na-selective microelectrodes; muscle strips at 0.5 Hz [6
8.5–30 Na-selective microelectrodes; myocytes at rest [80
5.1–21 SBFI-loaded myocytes at rest [9,10,79,81
17.5 23Na NMR; isolated perfused arrested hearts [82
Mouse 11.6 23Na NMR; isolated perfused heart [51
Mouse 14 SBFI-loaded myocytes at rest [2
Mouse LVH 23 SBFI-loaded myocytes at rest [2

Fine control of ATP-generating pathways in mitochondria and cytosol are critical to meet the energy demands of cardiac muscle. Supply must be matched to demand as failure to provide an adequate amount of ATP causes a decrease in cellular free energy leading to mechanical failure. The heart utilizes more energy than any other organ — with 2% of its total ATP reserves consumed per beat, it turns over its total ATP pool in less than 1 min and utilizes 6 kg of ATP every day [2123]. This enormous energy demand is related primarily to ATP-dependent processes driving EC coupling [24]. About 70–75% of total intracellular ATP is used for force generation powering work output, with the remaining 25–30% is used for basal metabolism [2527]. In terms of force generation, it is estimated that the actomyosin ATPase accounts for 76%, SERCA (sarcoendoplasmic reticulum Ca2+ ATPase) 15% and NKA for 9% of ATP utilization [27].

To synthesize the ATP required for normal function, the adult heart converts chemical energy primarily stored in free fatty acids (FFAs) (60–90%) and pyruvate (derived from glucose and lactate 10–40%) into mechanical energy for contraction [28]. The delivery of metabolic substrates, their selection, uptake and oxidation to generate acetyl-CoA for tricarboxylic acid (TCA) cycle entry and ATP generation in the electron transport chain (ETC) comprises three stages of myocardial ATP supply as summarized in Figure 1. However, cardiac workload varies constantly, including several-fold increase in cardiac output during exercise, thus requiring rapid and continuous matching of ATP supply to demand. This renders the heart a metabolic omnivore, giving it a high degree of substrate flexibility to rapidly switch substrate preference and utilization [28]. The apparent opposing relationship between carbohydrates and FFAs in the heart is, in part, due to the Randle (glucose–fatty acid) cycle, thus optimizing energy supply by avoiding energetic inefficiency and ‘waste’ [29].

First identified in the early 20th century, and now a well-established energy starvation hypothesis, it is proposed that maladaptive metabolic remodelling precedes, initiates and maintains adverse contractile dysfunction in hypertrophy and HF [23,24]. Advances in analytical technologies and understanding of metabolic mechanisms have improved our insights into the phenomenon and helped to classify metabolic alterations leading to myocardial energy starvation into those related to substrate utilization, intermediary metabolism and energetics. Using in vivo31P nuclear magnetic resonance (NMR), Neubauer [23] found that the myocardial phosphocreatine-to-ATP ratio (PCr:ATP) can be used as a reliable prognostic indicator of dilated cardiomyopathy (DCM) where 44% of DCM patients with a PCr:ATP of <1.6 died of cardiovascular causes vs. 5% with a PCr:ATP of >1.6. Cardiac hypertrophy induces a switch in substrate utilization from dominant FFA oxidation towards carbohydrate utilization which is similar to the foetal metabolic phenotype [24,3032]. The onset of this switch (and thereby the stage at which it could potentially be targeted therapeutically) is currently debated as numerous studies suggest that ATP levels are sustained during the early stages of remodelling and only decrease (30–40%) during advanced stages of HF [3337]. There have also been numerous preclinical studies as well as clinical data inferring mitochondrial respiratory impairment (complex activities and/or altered expression of the ETC complexes, ATP synthase and adenine nucleotide translocase) in hypertrophy and HF [3840].

EC coupling, specifically Ca handling by SERCA, has also been linked to the time course of metabolic alterations during hypertrophy: SERCA preferentially uses glycolytically derived ATP over OXPHOS (oxidative phosphorylation) [41] and therefore switching to a more glycolytic phenotype during hypertrophy, and HF could reflect increased SERCA activity to sustain adequate Ca homeostasis. NKA pump also requires glycolysis for normal Na homeostasis, potentially due to preferential fuelling of NKA by cytosolic glycolytically derived ATP and its spatial proximity to the pump [4,42,43]. However, the substrate switch and energetic deficit alone cannot explain Na accumulation observed in hypertrophy and failure. The debate is similar to the arguments about Na elevation in ischaemia and revolves around energetic inhibition of the NKA: the substrate switch from fatty acids to glucose leads to impaired energetic reserve and decline in cytosolic ATP, thus limiting the energy supply to the pump leading to Na accumulation. However, it has been previously shown that even during severe metabolic stress such as ischaemia, intracellular Na rises at a time when the total ATP concentration greatly exceeds the Km for the pump (∼0.1–0.8 mmol/l) and the free energy of ATP exceeds that required for pump activity (∼44 kJ/mol) [44].

In spite of significant evidence to support the concomitance of Nai overload and metabolic remodelling during cardiac hypertrophy and HF, there have been very few studies investigating the interaction between these pathophysiological events. Using isolated rat mitochondria, Iwai et al. [45] demonstrated that increasing extramitochondrial Na (Naex) from physiological (12.5 mM) to supraphysiological (≥25 mM) concentrations significantly reduced state 3 respiration, suggesting reduced mitochondrial ATP supply as well as reduced mitochondrial membrane potential. However, the present study offered no insights into the mechanism underlying the effect of Nai overload on whole cell metabolism.

A series of studies focusing on the mitochondrial transport of Na and Ca and its relationship with mitochondrial ATP production showed a stimulation of mitochondrial ATP production by Ca; however, the mitochondrial Ca transport kinetics and its regulation by Nai are still not completely understood [4548]. The majority of Camito uptake is by the Camito uniporter (MCU), while the Na/Camito exchanger (NCLX) is thought to be the predominant mechanism for Ca extrusion [49] (Figure 1). The impact of Nai on Cam has been elucidated by Cox and Matlib [46] using fura-2 to measure Cam in isolated cardiac mitochondria from healthy rabbits. Mitochondria incubated with increasing concentrations of Naex using NaCl in the physiological range showed reduced Camito as well as reduced NADH production and state 3 respiration. On the other hand, inhibition of NCLX with three inhibitors (from highest to lowest potency: CGP-37157 > clonazepam > d-cis-diltizam) and the MCU inhibitor ruthenium red substantially increased Cam, NADH production and state 3 respiration in a dose-dependent manner. This study supported the findings of Iwai et al. [45] and the hypothesis that Nai overload dysregulates ATP supply–demand matching.

However, this study did not provide information on the beat-to-beat kinetics of Cam transport and its relation to mitochondrial energy production.

Isolated mitochondria experiments should be treated with caution, given the measurements are performed in the absence of important ATP sinks (myosin ATPase, NKA and SERCA) and substrate utilization pathways (glycolysis and β-oxidation). More recently, Maack et al. [50] used isolated guinea pig cardiomyocytes to measure Cai and Cam during systole and diastole as well as NADH, thereby providing insights into the beat-to-beat regulation of Cam during increased Nai. This study showed that both systolic and diastolic Cam are significantly reduced by Nai elevation. Correspondingly, the percentage of NAD(H) in the reduced form was maintained at ∼62% in the control group, but was significantly lower in the high Nai group. In spite of these changes in Ca and [NADH], Nai elevation did not affect the mitochondrial membrane potential (ΔΨm). Furthermore, NCLX inhibition by CGP-37157 was shown to significantly elevate diastolic Cam. As these effects were not altered by ruthenium red inhibition of the MCU, it is likely a consequence of increased Cam extrusion via NCLX on a beat-to-beat basis. The outcome of this study further supported the argument that Nai is an important regulator of cardiac bioenergetics. However, it remains unclear whether this is truly reflective of a regulatory mechanism in the beating heart and, if so, which metabolic pathways are most affected by Nai overload.

The studies examining the impact of Nai on mitochondrial ATP provision published to date are subject to major experimental caveats, thus making direct mechanistic translation to in situ perfused and in vivo myocardium difficult. Specifically, these studies lack integrated experimental approaches as they have been limited to isolated organelles and cells at subphysiological temperatures with limited metabolic readouts.

To elucidate the importance of the link between Nai and ATP supply–demand matching in the beating heart, experimental models are required either ex vivo or in vivo where the heart is perfused under physiologically relevant conditions and where Nai elevation can be induced and reliably measured. To elucidate concomitant changes in substrate metabolism or energetics, it is also necessary to be able to quantify a wide range of metabolites involved in energy homeostasis in these models. We have previously applied and validated techniques able to measure intracellular Nai in the Langendorff perfused mouse [51,52] or rat heart preparations [53] using NMR.

23Na NMR has historically been used to distinguish the small intra Nai versus large extracellular Nae pools employing paramagnetic shift reagents such as Tm(DOTP) [thulium (III) 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonate)] [51] to separate the two. However, these reagents are efficient chelators of Ca and Mg leading to modified ion homeostasis and reduced cardiac contractility [51]. As a result, shift reagents exhibit significant toxicity precluding their use in vivo and question their validity for measuring Naiex vivo. In contrast, multiple quantum-filtered 23Na NMR, which exploits the quadrupolar property of the 23Na nucleus, has shown potential to probe intra and extracellular pools of Na in the absence of shift reagent and therefore under more physiological conditions [54,55]. We have previously investigated the use of these techniques in the perfused mouse heart where we were able to measure elevated intracellular Na in response to the cardiac glycoside ouabain as well as in response to modified buffer compositions, for example, in the absence of K, Ca or Mg. We were further able to verify previous studies showing that the PLM3SA mouse has a chronic elevation of basal Nai compared with wild-type hearts. Crucially, NMR is also able to probe cardiac energetics by 31P NMR in the same hearts where it is possible to measure the concentrations of ATP, PCr, Pi, intracellular pH as well as PCr:ATP ratio and thereby derive estimates of the Gibb's free energy. Newly emerging techniques such as metabolomics also enable end-point measurements of metabolites in snap frozen extracted myocardial tissue and coronary effluent using either high-resolution NMR or mass spectrometry [53,56,57].

Figure 3 shows the example NMR spectra of two different Langendorff perfused mouse hearts acquired using our previously reported NMR protocols. The left-hand panel displays data from a wild-type control mouse heart with normal baseline Nai, while the right-hand panel displays data from a hypertrophic mouse heart subject to aortic constriction [2]. The dry weight of the control heart was 30 mg while that of the banded heart was 58 mg measured at the end of the experiment. Figure 3a,b shows the 31P NMR spectra acquired with baseline function. Figure 3c,d shows the triple-quantum-filtered 23Na NMR spectra acquired with baseline function. Figure 3e,f shows the conventional single-quantum-filtered 23Na NMR spectra acquired at the end of the experiment following infusion of 5 mM Tm(DOTP) to shift the large extracellular Nae signal and enable quantification of the small intracellular Nai signal. Our previous work suggested that the TQF signal in Figure 3c,d consists of a contribution both from the intracellular and extracellular pools of Na, but that the large bulk isotropic signal from the buffer is largely suppressed. These experiments highlight the ability of such NMR techniques to probe both cardiac energetics using 31P NMR and Nai using triple-quantum-filtered 23Na NMR in the same preparation [51]. The data presented here also highlight experimental challenges in quantifying Nai in these hearts. Total myocardial Nai is clearly elevated under conditions of hypertrophy; however, so too is the tissue mass and intracellular volume [51]. Absolute quantification of such data is subject to many experimental assumptions including a phenomenological scaling factor for the NMR observability of Nai and a scaling factor to estimate intracellular volume. Despite obvious limitations in the methodology, NMR offers unique insights into Na ion homeostasis and cardiac energetics under both physiological and pathophysiological conditions. Additionally, there has resurgence in interest applying MRI techniques for imaging Na distribution in vivo [58]. 23Na is the second most sensitive nucleus for in vivo detection by NMR after 1H; however, sensitivity and spatial resolution remain an issue as well as the ability to separate intra- versus extracellular pools of Na which is also challenging.

Representative 31P NMR spectra, triple-quantum-filtered 23Na and conventional 1D 23Na NMR spectra from perfused control and hypertrophied mouse hearts.

Figure 3.
Representative 31P NMR spectra, triple-quantum-filtered 23Na and conventional 1D 23Na NMR spectra from perfused control and hypertrophied mouse hearts.

The spectra displayed in the left panel (a, c and e) are from a control heart, while those displayed in the right panel (b, d and f) are from a hypertrophied heart. All NMR data were acquired as previously described [51] using a Bruker Avance III 400 MHz wide-bore spectrometer. Briefly, a and b show 31P spectra, c and d show triple-quantum-filtered (TQF) 23Na NMR spectra, while e and f show conventional single-quantum 23Na NMR spectra acquired at the end of the perfusion during infusion of 5 mM Tm(DOTP).

Figure 3.
Representative 31P NMR spectra, triple-quantum-filtered 23Na and conventional 1D 23Na NMR spectra from perfused control and hypertrophied mouse hearts.

The spectra displayed in the left panel (a, c and e) are from a control heart, while those displayed in the right panel (b, d and f) are from a hypertrophied heart. All NMR data were acquired as previously described [51] using a Bruker Avance III 400 MHz wide-bore spectrometer. Briefly, a and b show 31P spectra, c and d show triple-quantum-filtered (TQF) 23Na NMR spectra, while e and f show conventional single-quantum 23Na NMR spectra acquired at the end of the perfusion during infusion of 5 mM Tm(DOTP).

Close modal

Nai has inadvertently been a known therapeutic target in HF for the last 200 years, and the established example of in vivo use of Nai modulation is the administration of cardiac glycosides (such as digoxin) which are potent inhibitors of NKA. Cardiac glycosides elevate Nai and lead to a positive inotropic response (due to release of Ca) that can vary considerably between species. They are steroidal-like compounds found endogenously under normal conditions (e.g. ouabain, digoxin and bufalin) and are elevated in patients with renal failure [59] and HF [60]. However, their clinical use for the treatment of HF is a cautionary tale and limited due to their energetically costly as well as pro-arrhythmic properties [61]. Nevertheless, they remain useful tool for elevating Nai in experimental models [5153,56,62]. In spite of the substantial in vitro and preclinical evidence to support the targeting of the substrate switch therapeutically, there has been limited successes that have been translated into the clinic. For example, sodium dichloroacetate (pyruvate dehydrogenase kinase inhibitor) appeared to improve contractile performance in 10 HF patients, but a vehicle control group was not included in this study [63,64]. Trimetazadine is currently prescribed for longer term inhibition of FFA oxidation and has been shown to reduce angina and improve cardiac function in patients with DCM [65,66], although these improvements were modest. Given the limited clinical success of targeting substrate utilization to date, it is important to continue to evaluate the potential of targeting other aspects of cardiac metabolism, such as intermediary pathways leading to ATP supply. This could also help identify the role metabolic remodelling plays in transition from pathological hypertrophy towards HF. The question remains whether early prevention of myocardial Nai elevation could either prevent the origin or alter the course of metabolic derangement in pathological hypertrophy leading to energy starvation and cardiac death. This hypothesis warrants further study including the ongoing development of therapeutics that target these interconnected pathophysiological events.

Camito

mitochondrial calcium

DCM

dilated cardiomyopathy

EC

excitation–contraction

ETC

electron transport chain

FFA

free fatty acids

HF

heart failure

MCU

mitochondrial calcium uniporter

Naex

extramitochondrial sodium

Nai

intracellular sodium

NCLX

mitochondrial sodium–calcium exchanger

NCX

sarcolemmal sodium–calcium exchanger

NKA

sodium/potassium ATPase

NMR

nuclear magnetic resonance

OXPHOS

oxidative phosphorylation

PCr:ATP

phosphocreatine-to-ATP ratio

PLM

phospholemman

SBFI

sodium-binding benzofuran isophthalate

SERCA

sarcoendoplasmic reticulum Ca2+ ATPase

Tm(DOTP)

thulium (III) 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonate)

D.A., T.R.E., B.A.O.B. and M.J.S. conceived and designed research. D.A. drafted the manuscript. D.A. and T.R.E. prepared figures. D.A., B.A.O.B., T.R.E. and M.J.S. edited, revised the manuscript and approved the final version of the manuscript.

This work was supported by a British Heart Foundation Programme Grant [RG/12/4/29426] to M.J.S. and King's College London British Heart Foundation Centre of Research Excellence [RE/08/003]. The research was supported by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy's and St Thomas' NHS Foundation Trust and King's College London; the Centre of Excellence in Medical Engineering funded by the Wellcome Trust and EPSRC under grant number [WT 088641/Z/09/Z] and the King's College London and UCL Comprehensive Cancer Imaging Centre funded by the CRUK and EPSRC in association with the MRC and DoH (England). The views expressed are those of the author and not necessarily those of the NHS, the NIHR or the Department of Health.

The authors thank Anja Karlstaedt, Heinrich Taegtmeyer, David Sanchez-Tatay, Marina Basalay, Andrew Atkinson, Alpesh Thakker and Daniel Tennant for their contributions to the Nai metabolism studies.

The Authors declare that there are no competing interests associated with the manuscript.

1
Shattock
,
M.J.
(
2009
)
Phospholemman: its role in normal cardiac physiology and potential as a drugable target in disease
.
Curr. Opin. Pharmacol.
9
,
160
166
2
Boguslavskyi
,
A.
,
Pavlovic
,
D.
,
Aughton
,
K.
,
Clark
,
J.E.
,
Howie
,
J.
,
Fuller
,
W.
et al.
(
2014
)
Cardiac hypertrophy in mice expressing unphosphorylatable phospholemman
.
Cardiovasc. Res.
104
,
72
82
3
Okamoto
,
K.
,
Wang
,
W.
,
Rounds
,
J.
,
Chambers
,
E.A.
and
Jacobs
,
D.O.
(
2001
)
ATP from glycolysis is required for normal sodium homeostasis in resting fast-twitch rodent skeletal muscle
.
Am. J. Physiol. Endocrinol. Metab.
281
,
E479
E488
4
Cross
,
H.R.
,
Radda
,
G.K.
and
Clarke
,
K.
(
1995
)
The role of Na+/K+ ATPase activity during low flow ischemia in preventing myocardial injury: a 31P, 23Na and 87Rb NMR spectroscopic study
.
Magn. Reson. Med.
34
,
673
685
5
Murphy
,
E.
and
Eisner
,
D.A.
(
2009
)
Regulation of intracellular and mitochondrial sodium in health and disease
.
Circ. Res.
104
,
292
303
6
Shattock
,
M.J.
and
Bers
,
D.M.
(
1989
)
Rat vs. rabbit ventricle: Ca flux and intracellular Na assessed by ion-selective microelectrodes
.
Am. J. Physiol.
256
,
C813
C822
7
Kohlhaas
,
M.
,
Liu
,
T.
,
Knopp
,
A.
,
Zeller
,
T.
,
Ong
,
M.F.
,
Bohm
,
M.
et al.
(
2010
)
Elevated cytosolic Na+ increases mitochondrial formation of reactive oxygen species in failing cardiac myocytes
.
Circulation
121
,
1606
1613
8
Liu
,
T.
and
O'Rourke
,
B.
(
2008
)
Enhancing mitochondrial Ca2+ uptake in myocytes from failing hearts restores energy supply and demand matching
.
Circ. Res.
103
,
279
288
9
Harrison
,
S.M.
,
McCall
,
E.
and
Boyett
,
M.R.
(
1992
)
The relationship between contraction and intracellular sodium in rat and guinea-pig ventricular myocytes
.
J. Physiol.
449
,
517
550
10
Despa
,
S.
,
Islam
,
M.A.
,
Weber
,
C.R.
,
Pogwizd
,
S.M.
and
Bers
,
D.M.
(
2002
)
Intracellular Na+ concentration is elevated in heart failure but Na/K pump function is unchanged
.
Circulation
105
,
2543
2548
11
Neubauer
,
S.
,
Newell
,
J.B.
and
Ingwall
,
J.S.
(
1992
)
Metabolic consequences and predictability of ventricular fibrillation in hypoxia. A 31P- and 23Na-nuclear magnetic resonance study of the isolated rat heart
.
Circulation
86
,
302
310
12
Pogwizd
,
S.M.
,
Sipido
,
K.R.
,
Verdonck
,
F.
and
Bers
,
D.M.
(
2003
)
Intracellular Na in animal models of hypertrophy and heart failure: contractile function and arrhythmogenesis
.
Cardiovasc. Res.
57
,
887
896
13
Verdonck
,
F.
,
Volders
,
P.G.
,
Vos
,
M.A.
and
Sipido
,
K.R.
(
2003
)
Intracellular Na+ and altered Na+ transport mechanisms in cardiac hypertrophy and failure
.
J. Mol. Cell. Cardiol.
35
,
5
25
14
Fuller
,
W.
,
Tulloch
,
L.B.
,
Shattock
,
M.J.
,
Calaghan
,
S.C.
,
Howie
,
J.
and
Wypijewski
,
K.J.
(
2013
)
Regulation of the cardiac sodium pump
.
Cell. Mol. Life Sci.
70
,
1357
1380
15
Pavlovic
,
D.
,
Fuller
,
W.
and
Shattock
,
M.J.
(
2013
)
Novel regulation of cardiac Na pump via phospholemman
.
J. Mol. Cell. Cardiol.
61
,
83
93
16
Shattock
,
M.J.
,
Ottolia
,
M.
,
Bers
,
D.M.
,
Blaustein
,
M.P.
,
Boguslavskyi
,
A.
,
Bossuyt
,
J.
et al.
(
2015
)
Na+/Ca2+ exchange and Na+/K+-ATPase in the heart
.
J. Physiol.
593
,
1361
1382
17
Pieske
,
B.
,
Maier
,
L.S.
,
Piacentino
, III,
V.
,
Weisser
,
J.
,
Hasenfuss
,
G.
and
Houser
,
S.
(
2002
)
Rate dependence of [Na+]i and contractility in nonfailing and failing human myocardium
.
Circulation
106
,
447
453
18
Verdonck
,
F.
,
Volders
,
P.G.
,
Vos
,
M.A.
and
Sipido
,
K.R.
(
2003
)
Increased Na+ concentration and altered Na/K pump activity in hypertrophied canine ventricular cells
.
Cardiovasc. Res.
57
,
1035
1043
19
Bossuyt
,
J.
,
Ai
,
X.
,
Moorman
,
J.R.
,
Pogwizd
,
S.M.
and
Bers
,
D.M.
(
2005
)
Expression and phosphorylation of the Na-pump regulatory subunit phospholemman in heart failure
.
Circ. Res.
97
,
558
565
20
Murphy
,
E.
and
Allen
,
D.G.
(
2009
)
Why did the NHE inhibitor clinical trials fail?
J. Mol. Cell. Cardiol.
46
,
137
141
21
Balaban
,
R.S.
(
2002
)
Cardiac energy metabolism homeostasis: role of cytosolic calcium
.
J. Mol. Cell. Cardiol.
34
,
1259
1271
22
Mootha
,
V.K.
,
Arai
,
A.E.
and
Balaban
,
R.S.
(
1997
)
Maximum oxidative phosphorylation capacity of the mammalian heart
.
Am. J. Physiol.
272
,
H769
H775
PMID:
[PubMed]
23
Neubauer
,
S.
(
2007
)
The failing heart — an engine out of fuel
.
N. Engl. J. Med.
356
,
1140
1151
24
Peterzan
,
M.A.
,
Lygate
,
C.A.
,
Neubauer
,
S.
and
Rider
,
O.J.
(
2017
)
Metabolic remodeling in hypertrophied and failing myocardium: a review
.
Am. J. Physiol. Heart Circ. Physiol.
313
,
H597
H616
25
Gibbs
,
C.L.
and
Loiselle
,
D.S.
(
2001
)
Cardiac basal metabolism
.
Jpn J. Physiol.
51
,
399
426
26
Gibbs
,
C.L.
(
2003
)
Cardiac energetics: sense and nonsense
.
Clin. Exp. Pharmacol. Physiol.
30
,
598
603
27
Schramm
,
M.
,
Klieber
,
H.G.
and
Daut
,
J.
(
1994
)
The energy expenditure of actomyosin-ATPase, Ca(2+)-ATPase and Na+,K(+)-ATPase in guinea-pig cardiac ventricular muscle
.
J. Physiol.
481
(
Pt 3
),
647
662
28
Stanley
,
W.C.
,
Recchia
,
F.A.
and
Lopaschuk
,
G.D.
(
2005
)
Myocardial substrate metabolism in the normal and failing heart
.
Physiol. Rev.
85
,
1093
1129
29
Hue
,
L.
and
Taegtmeyer
,
H.
(
2009
)
The Randle cycle revisited: a new head for an old hat
.
Am. J. Physiol. Endocrinol. Metab.
297
,
E578
E591
30
Aksentijević
,
D.
,
McAndrew
,
D.J.
,
Karlstädt
,
A.
,
Zervou
,
S.
,
Sebag-Montefiore
,
L.
,
Cross
,
R.
et al.
(
2014
)
Cardiac dysfunction and peri-weaning mortality in malonyl-coenzyme A decarboxylase (MCD) knockout mice as a consequence of restricting substrate plasticity
.
J. Mol. Cell. Cardiol.
75
,
76
87
31
Kolwicz
,
S.C.
and
Tian
,
R.
(
2011
)
Glucose metabolism and cardiac hypertrophy
.
Cardiovasc. Res.
90
,
194
201
32
Ventura-Clapier
,
R.
,
Garnier
,
A.
and
Veksler
,
V.
(
2004
)
Energy metabolism in heart failure
.
J. Physiol.
555
,
1
13
33
Akki
,
A.
and
Seymour
,
A.-M.L.
(
2009
)
Western diet impairs metabolic remodelling and contractile efficiency in cardiac hypertrophy
.
Cardiovasc. Res.
81
,
610
617
34
Chandler
,
M.P.
,
Kerner
,
J.
,
Huang
,
H.
,
Vazquez
,
E.
,
Reszko
,
A.
,
Martini
,
W.Z.
et al.
(
2004
)
Moderate severity heart failure does not involve a downregulation of myocardial fatty acid oxidation
.
Am. J. Physiol. Heart Circ. Physiol.
287
,
H1538
H1543
35
Starling
,
R.C.
,
Hammer
,
D.F.
and
Altschuld
,
R.A.
(
1998
)
Human myocardial ATP content and in vivo contractile function
.
Mol. Cell. Biochem.
180
,
171
177
36
Luptak
,
I.
,
Sverdlov
,
A.L.
,
Panagia
,
M.
,
Qin
,
F.
,
Pimentel
,
D.R.
,
Croteau
,
D.
et al.
(
2018
)
Decreased ATP production and myocardial contractile reserve in metabolic heart disease
.
J. Mol. Cell. Cardiol.
116
,
106
114
37
Beer
,
M.
,
Seyfarth
,
T.
,
Sandstede
,
J.
,
Landschütz
,
W.
,
Lipke
,
C.
,
Köstler
,
H.
et al.
(
2002
)
Absolute concentrations of high-energy phosphate metabolites in normal, hypertrophied, and failing human myocardium measured noninvasively with 31P-SLOOP magnetic resonance spectroscopy
.
J. Am. Coll. Cardiol.
40
,
1267
1274
38
Garnier
,
A.
,
Fortin
,
D.
,
Deloménie
,
C.
,
Momken
,
I.
,
Veksler
,
V.
and
Ventura-Clapier
,
R.
(
2003
)
Depressed mitochondrial transcription factors and oxidative capacity in rat failing cardiac and skeletal muscles
.
J. Physiol.
551
,
491
501
39
Sharov
,
V.G.
,
Todor
,
A.V.
,
Silverman
,
N.
,
Goldstein
,
S.
and
Sabbah
,
H.N.
(
2000
)
Abnormal mitochondrial respiration in failed human myocardium
.
J. Mol. Cell. Cardiol.
32
,
2361
2367
40
Lemieux
,
H.
,
Semsroth
,
S.
,
Antretter
,
H.
,
Höfer
,
D.
and
Gnaiger
,
E.
(
2011
)
Mitochondrial respiratory control and early defects of oxidative phosphorylation in the failing human heart
.
Int. J. Biochem. Cell Biol.
43
,
1729
1738
41
Xu
,
K.Y.
,
Zweier
,
J.L.
and
Becker
,
L.C.
(
1995
)
Functional coupling between glycolysis and sarcoplasmic reticulum Ca2+ transport
.
Circ. Res.
77
,
88
97
42
Dizon
,
J.
,
Burkhoff
,
D.
,
Tauskela
,
J.
,
Whang
,
J.
,
Cannon
,
P.
and
Katz
,
J.
(
1998
)
Metabolic inhibition in the perfused rat heart: evidence for glycolytic requirement for normal sodium homeostasis
.
Am. J. Physiol.
274
,
H1082
H1089
43
Weiss
,
J.
and
Hiltbrand
,
B.
(
1985
)
Functional compartmentation of glycolytic versus oxidative metabolism in isolated rabbit heart
.
J. Clin. Invest.
75
,
436
447
44
Fuller
,
W.
,
Parmar
,
V.
,
Eaton
,
P.
,
Bell
,
J.R.
and
Shattock
,
M.J.
(
2003
)
Cardiac ischemia causes inhibition of the Na/K ATPase by a labile cytosolic compound whose production is linked to oxidant stress
.
Cardiovasc. Res.
57
,
1044
1051
45
Iwai
,
T.
,
Tanonaka
,
K.
,
Inoue
,
R.
,
Kasahara
,
S.
,
Motegi
,
K.
,
Nagaya
,
S.
et al.
(
2002
)
Sodium accumulation during ischemia induces mitochondrial damage in perfused rat hearts
.
Cardiovasc. Res.
55
,
141
149
46
Cox
,
D.A.
and
Matlib
,
M.A.
(
1993
)
Modulation of intramitochondrial free Ca2+ concentration by antagonists of Na+-Ca2+ exchange
.
Trends Pharmacol. Sci.
14
,
408
413
47
Denton
,
R.M.
and
McCormack
,
J.G.
(
1990
)
Ca2+ as a second messenger within mitochondria of the heart and other tissues
.
Annu. Rev. Physiol.
52
,
451
466
48
Eisner
,
D.A.
,
Caldwell
,
J.L.
,
Kistamás
,
K.
and
Trafford
,
A.W.
(
2017
)
Calcium and excitation–contraction coupling in the heart
.
Circ. Res.
121
,
181
195
49
Hüser
,
J.
,
Blatter
,
L.A.
and
Sheu
,
S.-S.
(
2000
)
Mitochondrial calcium in heart cells: beat-to-beat oscillations or slow integration of cytosolic transients?
J. Bioenerg. Biomembr.
32
,
27
33
50
Maack
,
C.
,
Cortassa
,
S.
,
Aon
,
M.A.
,
Ganesan
,
A.N.
,
Liu
,
T.
and
O'Rourke
,
B.
(
2006
)
Elevated cytosolic Na+ decreases mitochondrial Ca2+ uptake during excitation-contraction coupling and impairs energetic adaptation in cardiac myocytes
.
Circ. Res.
99
,
172
182
51
Eykyn
,
T.R.
,
Aksentijević
,
D.
,
Aughton
,
K.L.
,
Southworth
,
R.
,
Fuller
,
W.
and
Shattock
,
M.J.
(
2015
)
Multiple quantum filtered 23Na NMR in the Langendorff perfused mouse heart: ratio of triple/double quantum filtered signals correlates with [Na]i
.
J. Mol. Cell. Cardiol.
86
,
95
101
52
Aksentijević
,
D.
,
Karlstäedt
,
A.
,
Basalay
,
M.
,
O'Brien
,
B.A.
,
Thakker
,
A.
,
Tennant
,
D.
et al.
(
2018
)
Causal link between intracellular Na overload and metabolic remodeling in the heart: uncoupling ATP supply and demand?
Heart
104
,
A2
A3
53
O'Brien
,
B.A.
,
Aksentijevic
,
D.
,
Eykyn
,
T.R.
and
Shattock
,
M.J.
(
2015
)
Ouabain induces metabolic changes in the absence of contractile work in the blebbistatin-perfused rat heart
.
J. Mol. Cell. Cardiol.
86
,
S47
54
Payne
,
G.S.
,
Seymour
,
A.-M.L.
,
Styles
,
P.
and
Radda
,
G.K.
(
1990
)
Multiple quantum filtered 23Na NMR spectroscopy in the perfused heart
.
NMR Biomed.
3
,
139
146
55
Dizon
,
J.M.
,
Tauskela
,
J.S.
,
Wise
,
D.
,
Burkhoff
,
D.
,
Cannon
,
P.J.
and
Katz
,
J.
(
1996
)
Evaluation of triple-quantum-filtered 23Na NMR in monitoring of Intracellular Na content in the perfused rat heart: comparison of intra- and extracellular transverse relaxation and spectral amplitudes
.
Magn. Reson. Med.
35
,
336
345
56
Aksentijevic
,
D.
,
Eykyn
,
T.R.
,
Fuller
,
W.
and
Shattock
,
M.J.
(
2015
)
Metabolic consequences of the acute and chronic myocardial sodium elevation
.
J. Mol. Cell. Cardiol.
86
,
1
78
57
Mansor
,
L.S.
,
Mehta
,
K.
,
Aksentijevic
,
D.
,
Carr
,
C.A.
,
Lund
,
T.
,
Cole
,
M.A.
et al.
(
2016
)
Increased oxidative metabolism following hypoxia in the type 2 diabetic heart, despite normal hypoxia signalling and metabolic adaptation
.
J. Physiol.
594
,
307
320
58
Madelin
,
G.
,
Lee
,
J.S.
,
Regatte
,
R.R.
and
Jerschow
,
A.
(
2014
)
Sodium MRI: methods and applications
.
Prog. Nucl. Magn. Reson. Spectrosc.
79
,
14
47
59
Kolmakova
,
E.V.
,
Haller
,
S.T.
,
Kennedy
,
D.J.
,
Isachkina
,
A.N.
,
Budny
,
G.V.
,
Frolova
,
E.V.
et al.
(
2011
)
Endogenous cardiotonic steroids in chronic renal failure
.
Nephrol. Dial. Transplant.
26
,
2912
2919
60
Kennedy
,
D.J.
,
Shrestha
,
K.
,
Sheehey
,
B.
,
Li
,
X.S.
,
Guggilam
,
A.
,
Wu
,
Y.
et al.
(
2015
)
Elevated plasma marinobufagenin, an endogenous cardiotonic steroid, is associated with right ventricular dysfunction and nitrative stress in heart failure
.
Circ. Heart Fail.
8
,
1068
1076
61
Gonano
,
L.A.
,
Sepulveda
,
M.
,
Rico
,
Y.
,
Kaetzel
,
M.
,
Valverde
,
C.A.
,
Dedman
,
J.
et al.
(
2011
)
Calcium-calmodulin kinase II mediates digitalis-induced arrhythmias
.
Circ. Arrhythm. Electrophysiol.
4
,
947
957
62
Matthews
,
P.M.
,
Williams
,
S.R.
,
Seymour
,
A.-M.
,
Schwartz
,
A.
,
Dube
,
G.
,
Gadian
,
D.G.
et al.
(
1982
)
A 31P-NMR study of some metabolic and functional effects of the inotropic agents epinephrine and ouabain, and the ionophore R02-2985 (X537A) in the isolated, perfused rat heart
.
Biochim. Biophys. Acta, Mol. Cell Res.
720
,
163
171
63
Bersin
,
R.M.
,
Wolfe
,
C.
,
Kwasman
,
M.
,
Lau
,
D.
,
Klinski
,
C.
,
Tanaka
,
K.
et al.
(
1994
)
Improved hemodynamic function and mechanical efficiency in congestive heart failure with sodium dichloroacetate
.
J. Am. Coll. Cardiol.
23
,
1617
1624
64
Bersin
,
R.M.
and
Stacpoole
,
P.W.
(
1997
)
Dichloroacetate as metabolic therapy for myocardial ischemia and failure
.
Am. Heart J.
134
,
841
855
65
Fragasso
,
G.
,
Palloshi
,
A.
,
Puccetti
,
P.
,
Silipigni
,
C.
,
Rossodivita
,
A.
,
Pala
,
M.
et al.
(
2006
)
A randomized clinical trial of trimetazidine, a partial free fatty acid oxidation inhibitor, in patients with heart failure
.
J. Am. Coll. Cardiol.
48
,
992
998
66
Tuunanen
,
H.
,
Engblom
,
E.
,
Naum
,
A.
,
Nagren
,
K.
,
Scheinin
,
M.
,
Hesse
,
B.
et al.
(
2008
)
Trimetazidine, a metabolic modulator, has cardiac and extracardiac benefits in idiopathic dilated cardiomyopathy
.
Circulation
118
,
1250
1258
67
Gray
,
R.P.
,
McIntyre
,
H.
,
Sheridan
,
D.S.
and
Fry
,
C.H.
(
2001
)
Intracellular sodium and contractile function in hypertrophied human and guinea-pig myocardium
.
Pflugers Arch.
442
,
117
123
68
Eisner
,
D.A.
,
Lederer
,
W.J.
and
Vaughan-Jones
,
R.D.
(
1984
)
The quantitative relationship between twitch tension and intracellular sodium activity in sheep cardiac Purkinje fibres
.
J. Physiol.
355
,
251
266
69
Lee
,
C.O.
,
Kang
,
D.H.
,
Sokol
,
J.H.
and
Lee
,
K.S.
(
1980
)
Relation between intracellular Na ion activity and tension of sheep cardiac Purkinje fibers exposed to dihydro-ouabain
.
Biophys. J.
29
,
315
330
70
Cohen
,
C.J.
,
Fozzard
,
H.A.
and
Sheu
,
S.S.
(
1982
)
Increase in intracellular sodium ion activity during stimulation in mammalian cardiac muscle
.
Circ. Res.
50
,
651
662
71
Lee
,
C.O.
and
Dagostino
,
M.
(
1982
)
Effect of strophanthidin on intracellular Na ion activity and twitch tension of constantly driven canine cardiac Purkinje fibers
.
Biophys. J.
40
,
185
198
72
Schmied
,
R.
,
Wang
,
G.X.
and
Korth
,
M.
(
1991
)
Intracellular Na+ activity and positive inotropic effect of sulmazole in guinea pig ventricular myocardium. Comparison with a cardioactive steroid
.
Circ. Res.
68
,
597
604
73
Wilde
,
A.A.
and
Kléber
,
A.G.
(
1986
)
The combined effects of hypoxia, high K+, and acidosis on the intracellular sodium activity and resting potential in guinea pig papillary muscle
.
Circ. Res.
58
,
249
256
74
Wang
,
G.X.
,
Schmied
,
R.
,
Ebner
,
F.
and
Korth
,
M.
(
1993
)
Intracellular sodium activity and its regulation in guinea-pig atrial myocardium
.
J. Physiol.
465
,
73
84
75
Jelicks
,
L.A.
and
Siri
,
F.M.
(
1995
)
Effects of hypertrophy and heart failure on [Na+]i in pressure-overloaded guinea pig heart
.
Am. J. Hypertens.
8
,
934
943
76
Liu
,
Y.
,
Cabo
,
C.
,
Salomonsz
,
R.
,
Delmar
,
M.
,
Davidenko
,
J.
and
Jalife
,
J.
(
1993
)
Effects of diacetyl monoxime on the electrical properties of sheep and guinea pig ventricular muscle
.
Cardiovasc. Res.
27
,
1991
1997
77
Liu
,
T.
,
Takimoto
,
E.
,
Dimaano
,
V.L.
,
DeMazumder
,
D.
,
Kettlewell
,
S.
,
Smith
,
G.
et al.
(
2014
)
Inhibiting mitochondrial Na+/Ca2+ exchange prevents sudden death in a guinea pig model of heart failure
.
Circ. Res.
115
,
44
54
78
Baudet
,
S.
,
Noireaud
,
J.
and
Léoty
,
C.
(
1991
)
Intracellular Na activity measurements in the control and hypertrophied heart of the ferret: an ion-sensitive micro-electrode study
.
Pflugers Arch.
418
,
313
318
79
Levi
,
A.J.
,
Lee
,
C.O.
and
Brooksby
,
P.
(
1994
)
Properties of the fluorescent sodium indicator ‘SBFI’ in rat and rabbit cardiac myocytes
.
J. Cardiovasc. Electrophysiol.
5
,
241
257
80
Grupp
,
I.
,
Im
,
W.B.
,
Lee
,
C.O.
,
Lee
,
S.W.
,
Pecker
,
M.S.
and
Schwartz
,
A.
(
1985
)
Relation of sodium pump inhibition to positive inotropy at low concentrations of ouabain in rat heart muscle
.
J. Physiol.
360
,
149
160
81
Donoso
,
P.
,
Mill
,
J.G.
,
O'Neill
,
S.C.
and
Eisner
,
D.A.
(
1992
)
Fluorescence measurements of cytoplasmic and mitochondrial sodium concentration in rat ventricular myocytes
.
J. Physiol.
448
,
493
509
82
Schepkin
,
V.D.
,
Choy
,
I.O.
,
Budinger
,
T.F.
,
Obayashi
,
D.Y.
,
Taylor
,
S.E.
,
DeCampli
,
W.M.
et al.
(
1998
)
Sodium TQF NMR and intracellular sodium in isolated crystalloid perfused rat heart
.
Magn. Reson. Med.
39
,
557
563
This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY).