Determination of sixteen nucleotides, nucleosides and bases using high-performance liquid chromatography and its application to the study of purine metabolism in hearts for transplantation

Smolenski, Ryszard T.; Lachno, D. Richard; Ledingham, Simon; Yacoub, M

doi:10.1016/s0378-4347(00)82125-8

Cited by 301 publications

(157 citation statements)

References 9 publications

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“…Next, serum components were extracted with 1.3 M perchloric acid (1:1 ratio). Levels of nucleotides were measured by a reverse phase-high pressure liquid chromatography (RP-HPLC) method using the LC system (Agilent Technologies 1100 series, USA), as described previously [15]. Results are presented as nmol/mg of dry tissue for heart samples and as µM for mouse serum.…”

Section: Measurement Of Nucleotides and Corresponding Levels Of Catabmentioning

confidence: 99%

An impaired metabolism of nucleotides underpins a novel mechanism of cardiac remodeling leading to Huntington's disease related cardiomyopathy

Toczek

Zielonka

Zukowska

et al. 2016

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Huntington's disease (HD) is mainly thought of as a neurological disease, but multiple epidemiological studies have demonstrated a number of cardiovascular events leading to heart failure in HD patients. Our recent studies showed an increased risk of heart contractile dysfunction and dilated cardiomyopathy in HD pre-clinical models. This could potentially involve metabolic remodeling, that is a typical feature of the failing heart, with reduced activities of high energy phosphate generating pathways. In this study, we sought to identify metabolic abnormalities leading to HD-related cardiomyopathy in pre-clinical and clinical settings. We found that HD mouse models developed a profound deterioration in cardiac energy equilibrium, despite AMP-activated protein kinase hyperphosphorylation. This was accompanied by a reduced glucose usage and a significant deregulation of genes involved in de novo purine biosynthesis, in conversion of adenine nucleotides, and in adenosine metabolism. Consequently, we observed increased levels of nucleotide catabolites such as inosine, hypoxanthine, xanthine and uric acid, in murine and human HD serum. These effects may be caused locally by mutant HTT, via gain or loss of function effects, or distally by a lack of trophic signals from central nerve stimulation. Either may lead to energy equilibrium imbalances in cardiac cells, with activation of nucleotide catabolism plus an inhibition of resynthesis. Our study suggests that future therapies should target cardiac mitochondrial dysfunction to ameliorate energetic dysfunction. Importantly, we describe the first set of biomarkers related to heart and skeletal muscle dysfunction in both pre-clinical and clinical HD settings.Keywords: Huntington's disease, cardiomyopathy, arrhythmia, energy imbalance, catabolism of nucleotides, heart failure 3 SUMMARY Huntington's disease (HD) is a fatal neurodegenerative disorder caused by a polyglutamine expansion in the huntingtin protein. HD-related cardiomyopathy has been widely described in different mouse models, however there is little known about the source of the pathological remodelling in HD hearts. We found that contractile dysfunction in HD settings might be caused by components of cellular energy imbalance, changes in catabolism of adenine nucleotides, steady-state internal redox derangements and an activation of AMPK, leading to a shift in the cardiac substrate preference. These changes were accompanied by increased concentrations of adenine nucleotide catabolites (inosine, hypoxanthine, xanthine and uric acid) and uridine in both HD mouse models and HD patients' plasma. These metabolites represent the first identified biomarkers related to striated muscle dysfunction in HD. Our study explores a mechanism that might lead to HD-related cardiomyopathy and opens new avenues for therapeutic treatments in HD. HIGHLIGHTS• Heart dysfunction in HD is caused by the altered metabolism of nucleotides in vivo • Altered energy imbalances may lead to heart malfunction in HD in vivo• Increased lev...

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Section: Measurement Of Nucleotides and Corresponding Levels Of Catabmentioning

confidence: 99%

An impaired metabolism of nucleotides underpins a novel mechanism of cardiac remodeling leading to Huntington's disease related cardiomyopathy

Toczek

Zielonka

Zukowska

et al. 2016

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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“…Separations were achieved on Hypersil BDS 125 x 3 mm, 3-mm particle size column (Agilent Technologies). Modifications were introduced into the original method (Smolenski et al, 1990). The mobile phase flowed at a rate of 0.5 mL/min and column temperature was 20.5ºC.…”

Section: Oxypurine and Purine Nucleoside Concentrations In Renal Veinmentioning

confidence: 99%

Ischemia-Reperfusion Injury in the Transplanted Kidney Based on Purine Metabolism Markers and Activity of the Antioxidant System

Domański¹,

Kłoda²,

Ciechanowski³

2012

Organ Donation and Transplantation - Public Policy and Clinical Perspectives

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“…The ATP, ADP, AMP NAD + , NADP + concentrations obtained in the blood were calculated according to factors given in the assay method (Smolenski et al 1990) and expressed as µΜ οf RBC. The concentrations of inosine, hypoxanthine, uric acid expressed as µΜ in the whole blood.…”

Section: Hplc Separation Of Purinementioning

confidence: 99%

Purine Turnover Metabolites and Selected Antioxidants in Blood of Goats during the Periparturient Period

et al. 2010

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The aim of this study was to examine the relationships between the erythrocyte concentrations of ATP, ADP, AMP, NAD + , NADP + and adenylate energy charge (AEC) and their metabolites: inosine (Ino), hypoxanthine (Hyp), uric acid (UA), selected blood antioxidants: superoxide dismutase (SOD), glutathione reductase (GR) of goats during the periparturient period.The study was conducted on 12 clinically healthy pregnant goats (Capra hircus) -75% genotype of the Polish noble white aged between 2-3 years. Blood was taken from the external jugular vein early in the morning before feeding and obtained twice (4 weeks and 1 week) before delivery and twice (2 and 3 weeks) after delivery.The ATP concentration did not change significantly. One week before delivery ADP, AMP and Ino concentrations were significantly higher and AEC value significantly lower compared to values in the third week after delivery (p ≤ 0.02, p ≤ 0.05, p ≤ 0.03 and p ≤ 0.01, respectively). One week before and two weeks after delivery we observed a significant increase in SOD and GR activities and an increase in NAD + and NADP + concentrations but a significant decrease in Hyp and UA concentrations. Constant ATP concentration and the changes in the dynamics and tendency of Hyp and UA concentrations show that in the periparturient period in goats, purine metabolism undergoes adaptive changes. The balance between resynthetic processes and adenine nucleotide degradation is retained in order to level the oxygen and energy lacks. The results of our study show that the maintenance of prooxidative homeostasis in goats of peripartal period depends mainly on enzymatic mechanisms.

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Determination of sixteen nucleotides, nucleosides and bases using high-performance liquid chromatography and its application to the study of purine metabolism in hearts for transplantation

Cited by 301 publications

References 9 publications

An impaired metabolism of nucleotides underpins a novel mechanism of cardiac remodeling leading to Huntington's disease related cardiomyopathy

An impaired metabolism of nucleotides underpins a novel mechanism of cardiac remodeling leading to Huntington's disease related cardiomyopathy

Ischemia-Reperfusion Injury in the Transplanted Kidney Based on Purine Metabolism Markers and Activity of the Antioxidant System

Purine Turnover Metabolites and Selected Antioxidants in Blood of Goats during the Periparturient Period

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