High performance liquid chromatography: a rapid isocratic method for determination of creatine compounds and adenine nucleotides in myocardial tissue

Sellevold, Olav F Münter; Jynge, Per; Aarstad, Kjell

doi:10.1016/s0022-2828(86)80917-8

Cited by 280 publications

(111 citation statements)

References 25 publications

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“…HEP data are summarized in Table 4. In normal animals, 31 P spectra recorded during the control period were characterized by high PCr/ ATP. Basal PCr/ATP was significantly decreased in hearts with LVH and further decreased in CHF hearts (Table 4, P Ͻ 0.05 vs. normal).…”

Section: Resultsmentioning

confidence: 99%

“…Spectra were obtained with the image-selected in vivo spectroscopy (ISIS) method, which defined a column 1.8 ϫ 1.8 cm 2 perpendicular to the LV wall. 31 P signal excitation was achieved with a 90°, adiabatic, B 1-insensitive, BIR-4 pulse. All chemical shifts were measured relative to the PCr peak, which was assigned a chemical shift of Ϫ2.55 ppm relative to 85% phosphonoacetic acid at 0 ppm.…”

mentioning

confidence: 99%

“…With the use of a forceps precooled to Ϫ70°C, at the end of the experiment, an epicardial biopsy was taken from five normal, four LVH, and four CHF ventricles for subsequent analysis of ATP and creatine content using HPLC techniques (31). The heart was then fixed in 10% buffered formalin.…”

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confidence: 99%

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Oxidative capacity in failing hearts

Gong

Liu

Liang

et al. 2003

American Journal of Physiology-Heart and Circulatory Physiology

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Although high-energy phosphate metabolism is abnormal in failing hearts [congestive heart failure (CHF)], it is unclear whether oxidative capacity is impaired. This study used the mitochondrial uncoupling agent 2,4-dinitrophenol (DNP) to determine whether reserve oxidative capacity exists during the high workload produced by catecholamine infusion in hypertrophied and failing hearts. Left ventricular hypertrophy (LVH) was produced by ascending aortic banding in 21 swine; 9 animals developed CHF. Basal myocardial phosphocreatine (PCr)/ATP measured with 31 P NMR spectroscopy was decreased in both LVH and CHF hearts (corresponding to an increase in free [ADP]), whereas ATP was decreased in hearts with CHF. Infusion of dobutamine and dopamine (each 20 g ⅐ kg Ϫ1 ⅐ min Ϫ1 iv) caused an approximate doubling of myocardial oxygen consumption (MV O2) in all groups and decreased PCr/ATP in the normal and LVH groups. During continuing catecholamine infusion, DNP (2-8 mg/kg iv) caused further increases of MV O2 in normal and LVH hearts with no change in PCr/ATP. In contrast, DNP caused no increase in MV O2 in the failing hearts; the associated decrease of PCr/ATP suggests that DNP decreased the mitochondrial proton gradient, thereby causing ADP to increase to maintain adequate ATP synthesis. heart failure; left ventricular hypertrophy; mitochondria; high-energy phosphates; nuclear magnetic resonance IN NORMAL MYOCARDIUM, it is unclear whether peak O 2 utilization is ultimately limited by maximal oxidative ATP synthetic capacity or by the maximal capacities of the myosin and other ATPases to utilize ATP. In failing myocardium, abnormalities of excitation-contraction coupling as well as downregulation of ␤-adrenergic receptors and the downstream adenylyl cyclase system (25) make it even more difficult to determine whether ATP synthetic capacity limits contractile performance. Hence, the hypothesis that primary "energy starvation" limits function in heart failure (14) remains to be rigorously tested in vivo despite evidence that left ventricular (LV) hypertrophy (LVH) and congestive heart failure (CHF) are associated with abnormalities of myocardial energy metabolism (2,21,22,38,39).Consequently, the present study was performed to determine whether administration of a classical mitochondrial uncoupling agent [2,4-dinitrophenol (DNP)] could further increase myocardial oxygen consumption (MV O 2 ) in hearts with compensated LVH or overt cardiac failure that were already functioning at a high work state produced by catecholamine stimulation. DNP accelerates intramitochondrial metabolism proximal to ATP synthase by decreasing the proton gradient across the inner mitochondrial membrane (17,30). In response to DNP, MV O 2 increases in concert with intermediary metabolism and electron transport activity to maintain the mitochondrial proton gradient that drives ATP synthesis (15,17,30). Although it is unlikely that DNP can define the maximal oxygen utilization capacity in the intact heart (8), it can be used to determine whether there ...

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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Oxidative capacity in failing hearts

Gong

Liu

Liang

et al. 2003

American Journal of Physiology-Heart and Circulatory Physiology

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“…Homogenization and deproteinization were performed in ice-cold 60% acetonitrile (v/v) using a Branson sonifier. Using two different high performance liquid chromatography protocols, the myocardial contents in ATP, ADP, and AMP, and the intermediary metabolites and end products of glycolysis were determined (17,18). The injury caused by the biopsies did not interfere with the triphenyltetrazolium chloride stainings.…”

Section: Methodsmentioning

confidence: 99%

Regulation of Glycolytic Flux in Ischemic Preconditioning

Vogt¹,

Poolman²,

Ackermann³

et al. 2002

Journal of Biological Chemistry

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Exact adjustment of the Embden-Meyerhof pathway (EMP) is an important issue in ischemic preconditioning(IP) because an attenuated ischemic lactate accumulation contributes to myocardial protection. However, precise mechanisms of glycolytic flux and its regulation in IP remain to be elucidated. In open chest pigs, IP was achieved by two cycles of 10-min coronary artery occlusion and 30-min reperfusion prior to a 45-min index ischemia and 120-min reperfusion. Myocardial contents in glycolytic intermediates were assessed by high performance liquid chromatographic analysis of serial myocardial biopsies under control conditions and IP. Detailed time courses of metabolite contents allow an in-depth description of EMP regulation during index ischemia using metabolic control analysis. IP reduced myocardial infarct size (control, 90.0 ؎ 3.1 versus 5.05 ؎ 2.1%; p < 0.001) and attenuated myocardial lactate accumulation (end-ischemic contents, 31.9 ؎ 4.47 versus 10.3 ؎ 1.26 mol/wet weight, p < 0.0001), whereby a decrease in anaerobic glycolytic flux by at least 70% could constantly be observed throughout index ischemia. By calculation of flux:metabolite co-responses, the mechanisms of glycolytic regulation were investigated. The continuous deceleration of EMP flux in control myocadium could neither be explained on the basis of substrate availability nor be attributed to regulatory "key enzymes," as multisite regulation was employed for flux adjustment. In myocardium subjected to IP, an even pronounced deceleration of EMP flux during index ischemia was observed. Again, the adjustment of EMP flux was because of multisite modulation without any evidence for flux limitation by substrate availability or a key enzyme. However, IP changed the regulatory properties of most EMP enzymes, and some of these patterns could not be explained on the basis of substrate kinetics. Instead, other regulatory mechanisms, which have previously not yet been described for EMP enzymes, must be considered. These altered biochemical properties of the EMP enzymes have not yet been described.Myocardial survival in states of supply/demand imbalance critically depends on cellular energy status (1). To limit energy deficit in conditions of energy shortage, e.g. hypoxia and ischemia, myocardial energy production switches from the preferential use of fatty acids to carbohydrates, thereby allowing maintenance of adequate ATP synthesis when decreased oxygen availability becomes the limiting factor (2-5). However, in zeroflow ischemia, experimental analysis has shown increased glycolysis to be a double-edged sword, as the accumulation of glycolytic end products (6, 7) outweighs the potential benefits of increased ATP synthesis.In accordance with this paradigm, myocardial protection by prior exposure to ischemic preconditioning (IP) 1 employs a limitation in myocardial energy deficit (8, 9). Because IP tremendously reduces ischemic myocardial energy demands (8), energy deficit is largely decreased at even reduced rates of anaerobic glycolytic ATP formation in ze...

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“…We examined the effect of resveratrol on the total adenine nucleotide content in Neuro2a ECP is normally maintained in the range of 70% to 100% in healthy cells and tissues that are provided with sufficient nutrition and oxygenation [35,36]. Thereupon, ECP can be seen as an indicator of "well-being" status of cells and organisms [37].…”

Section: Resveratrol Significantly Increases the Total Pool Of Adeninmentioning

confidence: 99%

Proenergetic effects of resveratrol in the murine neuronal cell line Neuro2a

Nguyen

Ooi

Piller

et al. 2013

Mol. Nutr. Food Res.

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Scope: Energy deficit is a common characteristic of neurodegenerative disorders, including Alzheimer's disease. Adenosine monophosphate activated protein kinase (AMPK) is a key enzyme maintaining energy balance by regulating the cellular uptake of glucose, β-oxidation of fatty acids, and expression of glucose transporter 4. Since resveratrol has been shown to increase the activity of AMPK, we hypothesized that it might influence energy metabolism in a model neuron-like cell line, murine Neuro2a cells.Methods and results: Resveratrol caused an elevation of adenosine triphosphate (ATP) and guanosine triphosphate (GTP) in a dose-dependent manner. The highest ATP and GTP levels achieved by treatment with resveratrol were 70.3 ± 8.2 nmol/mg protein (1.9-fold of control) and 27.2 ± 4.0 nmol/mg protein (1.7-fold of control), respectively, when cells were treated with 100 μM resveratrol for 6 h. Interestingly, increases in the total sum of all adenine nucleotides were found upon addition of resveratrol. Despite these increases in ATP, GTP, and the total adenine nucleotide pool, resveratrol treatment led to a decrease in glucose consumption and lactate release, suggesting that resveratrol does not increase energy production (e.g. via AMPK kinase activation) but rather inhibits energy-consuming processes.Conclusion: Resveratrol increases the levels of ATP and GTP, but without creating an additional glucose demand. treated with 100 µM resveratrol for 6 h. Interestingly, substantial increases in the total sum of all adenine nucleotides were found upon addition of resveratrol. Despite these increases in ATP, GTP and the total adenine nucleotide pool, resveratrol treatment led to a pronounced decrease in glucose consumption and lactate release, suggesting that resveratrol does not increase energy production (e.g. via AMPK kinase activation) but rather inhibits energy consuming processes.Conclusions: Resveratrol increases the levels of free high-energy nucleotides, including ATP and GTP, but without creating an additional glucose demand.4

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High performance liquid chromatography: a rapid isocratic method for determination of creatine compounds and adenine nucleotides in myocardial tissue

Cited by 280 publications

References 25 publications

Oxidative capacity in failing hearts

Oxidative capacity in failing hearts

Regulation of Glycolytic Flux in Ischemic Preconditioning

Proenergetic effects of resveratrol in the murine neuronal cell line Neuro2a

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