title: Accelerated degradation of adenine nucleotide in erythrocytes of patients with chronic renal failure.
abstract: Recently, we have shown that erythrocytes obtained from patients with chronic renal failure (CRF) exhibited an increased rate of ATP formation from adenine as a substrate. Thus, we concluded that this process was in part responsible for the increase of adenine nucleotide concentration in uremic erythrocytes. There cannot be excluded however, that a decreased rate of adenylate degradation is an additional mechanism responsible for the elevated ATP concentration. To test this hypothesis, in this paper we compared the rate of adenine nucleotide breakdown in the erythrocytes obtained from patients with CRF and from healthy subjects. Using HPLC technique, we evaluated: (1) hypoxanthine production by uremic RBC incubated in incubation medium: (a) pH 7.4 containing 1.2 mM phosphate (which mimics physiological conditions) and (b) pH 7.1 containing 2.4 mM phosphate (which mimics uremic conditions); (2) adenine nucleotide degradation (IMP, inosine, adenosine, hypoxanthine production) by uremic RBC incubated in the presence of iodoacetate (glycolysis inhibitor) and EHNA (adenosine deaminase inhibitor). The erythrocytes of healthy volunteers served as control. The obtained results indicate that adenine nucleotide catabolism measured as a hypoxanthine formation was much faster in erythrocytes of patients with CRF than in the cells of healthy subjects. This phenomenon was observed both in the erythrocytes incubated at pH 7.4 in the medium containing 1.2 mM inorganic phosphate and in the medium which mimics hyperphosphatemia (2.4 mM) and metabolic acidosis (pH 7.1). The experiments with EHNA indicated that adenine nucleotide degradation proceeded via AMP-IMP-Inosine-Hypoxanthine pathway in erythrocytes of both patients with CRF and healthy subjects. Iodoacetate caused a several fold stimulation of adenylate breakdown. Under these conditions: (a) the rate of AMP catabolites (IMP + inosine + adenosine + hypoxanthine) formation was substantially higher in the erythrocytes from patients with CRF; (b) in erythrocytes of healthy subjects degradation of AMP proceeded via IMP and via adenosine essentially at the same rate; (c) in erythrocytes of patients with CRF the rate of AMP degradation via IMP was about 2 fold greater than via adenosine. The results presented in this paper suggest that adenine nucleotide degradation is markedly accelerated in erythrocytes of patients with CRF.
authors.affiliation: Department of Biochemistry, Medical University of Gdańsk, Debinki, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Marlewski
authors.forename: R T
authors.initials: RT
authors.lastname: Smolenski
authors.forename: M
authors.initials: M
authors.lastname: Szolkiewicz
authors.forename: Z
authors.initials: Z
authors.lastname: Aleksandrowicz
authors.forename: B
authors.initials: B
authors.lastname: Rutkowski
authors.forename: J
authors.initials: J
authors.lastname: Swierczynski
mesh_terms: D000227:Adenine Nucleotides / Q000097:blood / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000097:blood / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000097:blood / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000097:blood / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D006801:Humans; D019271:Hypoxanthine / Q000097:blood; D007676:Kidney Failure, Chronic / Q000097:blood*; D008297:Male; D008875:Middle Aged
chemical_list: D000227:Adenine Nucleotides; D019271:Hypoxanthine; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate