Impaired l-arginine metabolism marks endothelial dysfunction in CD73-deficient mice

Abstract

Changes in the ecto-5′-nucleotidase activity—an extracellular nucleotide catabolic enzyme may lead to the inflammation and endothelial dysfunction. We investigated the effect of CD73 deletion on the endothelial function and l -arginine metabolism in various age groups of mice. 1-,3-,6-, and 12-month-old, male C57BL/6 J wild type (WT) and C57BL/6 J CD73−/− (CD73−/−) mice were used. Blood samples were used for the analysis of adenine nucleotide concentrations. Serum samples were analyzed for the concentration of amino acids, Interleukin 6 (IL-6), Intercellular Adhesion Molecule 1 (ICAM-1), Vascular Cell Adhesion Molecule 1 (VCAM-1), and endothelial nitric oxide synthase (eNOS) level. Serum and aortic nitrate/nitrite, as well as aortic arginase and NOS activity in endothelial cells (EC) were evaluated. CD73 deletion led to age-dependent increase in IL-6, ICAM-1, and VCAM-1 concentration compared to WT. All CD73−/− mice age groups were characterized by reduced l -Arginine concentration and eNOS level. Significantly lower NOS activity was noticed in EC isolated from CD73−/− mice lungs in comparison to EC isolated from WT lungs. The l -Arginine/ADMA ratio in the CD73−/− decreased in age-dependent manner in comparison to WT. The nitrate/nitrite ratio was reduced in serum and in aortas of 6-month-old CD73−/− mice as compared to WT. The ornithine/arginine and ornithine/citrulline ratios were increased in CD73−/− compared to controls. Blood (erythrocyte) Adenosine-5′-triphosphate and Adenosine-5′-diphosphate levels were reduced in favor to higher blood Adenosine-5′-monophosphate concentration in CD73−/− mice in comparison to WT. The CD73 deletion leads to the development of age-dependent endothelial dysfunction in mice, associated with impaired l -arginine metabolism. CD73 activity seems to protect endothelium. Electronic supplementary material The online version of this article (10.1007/s11010-019-03537-4) contains supplementary material, which is available to authorized users.

High Throughput Procedure for Comparative Analysis of In Vivo Cardiac Glucose or Amino Acids Use in Cardiovascular Pathologies and Pharmacological Treatments

“…CD73 knock-out results in a decrease in basal coronary flow, increased fibrosis, and greater cardiomyocyte hypertrophy after a transverse aortic constriction procedure [ 42 , 43 ]. Moreover, our earlier studies indicated that the deletion of CD73 leads to aortic valve and endothelial dysfunction [ 44 , 45 ]. It is well known that deterioration in heart and vascular function may result in changes in cardiac metabolism.…”

Macrophage-Derived Adenosine Deaminase 2 Correlates with M2 Macrophage Phenotype in Triple Negative Breast Cancer

“…The supernatant was brought to pH 6.0–6.5 using 3 M K3PO4 and analyzed with HPLC according to the method described earlier. [ 45 , 46 ] ADA1 activity was calculated from the difference between tADA and ADA2 activities.…”

“…The supernatant was brought to pH 6.0-6.5 using 3 M K3PO4 and analyzed with HPLC according to the method described earlier. [45,46] ADA1 activity was calculated from the difference between tADA and ADA2 activities. Concentration of soluble CD163 and galectin-3 in human plasma were measured using enzyme-linked immunosorbent assay kits according to the manufacturer's protocols.…”

Increased expression levels of inflammatory cytokines and adhesion molecules in lipopolysaccharide‑induced acute inflammatory apoM‑/‑ mice

“…To the best of the authors' knowledge, the direct upstream and downstream relationship between apoM and IL-10 has yet to be elucidated, thus, in the present study, the expression levels of IL-10 in apoM −/− mice were determined and further analyses were performed to confirm the interaction between IL-10 and apoM. Serum ICAM-1 and VCAM-1 are generally detected by enzyme-linked immunosorbent assay ( 36 , 37 ), but in the present study, they were not detected using western blotting and enzyme-linked immunosorbent assay, probably because their expression levels were too low.…”