Effect of estradiol on enzymes of vascular extracellular nucleotide metabolism

Abstract

Purpose: Estrogens have beneficial effects on the cardiovascular system, promoting vasodilation, endothelial cells growth, relaxation, and regulation of blood pressure. Some of these effects could be associated with the purinergic system known for the control of vasodilation, inflammation, and platelet function. The aim of our study was the evaluation of ATP, AMP, and adenosine extracellular catabolism, catalyzed by ectonucleoside triphosphate diphosphohydrolase-1 (CD39), ecto-5'-nucleotidase (CD73), and ecto-adenosine deaminase (eADA) in mouse aortas.

Methods: Extracellular hydrolysis of ATP, AMP, and adenosine was estimated on the aortic surface of 3-month-old female and male C57BL/6 J wild-type (WT) mice, in female WT mouse aortas incubated for 48 h in the presence or absence of 100 nM estradiol, and in WT female mouse and ApoE-/-LDL-R-/- aortas. The conversion of substrates to products was analyzed by high-pressure liquid chromatography (HPLC).

Results: We demonstrated significantly higher adenosine deamination rate in WT male vs. female mice (p = 0.041). We also noted the lower adenosine hydrolysis in aortas exposed to estradiol, as compared with the samples incubated in estradiol-free medium (p = 0.043). Finally, we observed that adenosine conversion to inosine was significantly higher on the surface of ApoE-/-LDL-R-/- aortas compared with WT mice (p = 0.001). No such effects were noted in ATP and AMP extracellular hydrolysis.

Conclusion: We conclude that estradiol inhibits the extracellular degradation of adenosine to inosine, which may be an element of its vascular protective effect, as it will lead to an increase in extracellular adenosine concentration. We can also assume that during the development of the atherosclerotic process, the protective role of estradiol in the regulation of adenosine degradation may be obscured by other pathogenic factors.

Keywords: Aorta; CD39; CD73; Endothelium; Estrogens; Mice; eADA.

Fig. 1

Comparison of eADA, CD73, and CD39 activities in WT female (F) (n = 6) and male (M) (n = 6) mice aortic endothelial cells. Data are presented as a boxplots

Fig. 2

Activities of ADA, CD73, and CD39 in WT female mice aortic endothelial cells after treatment with 100 nM estradiol (F/E+) (n = 6) vs. controls (F/E-) (n = 6). Data are presented as a boxplots

Fig. 3

Activities of ADA, CD73, and CD39 in WT (F control) (n = 11) and ApoE-/-LDL-R-/- (F LDL-/-ApoE-/-) (n = 8) female mice aortic endothelial cells. Data are presented as a boxplots