Regulation of Hypoxic–Adenosinergic Signaling by Estrogen: Implications for Microvascular Injury

Abstract

Loss of estrogen, as occurs with normal aging, leads to increased inflammation, pathologic angiogenesis, impaired mitochondrial function, and microvascular disease. While the influence of estrogens on purinergic pathways is largely unknown, extracellular adenosine, generated at high levels by CD39 and CD73, is known to be anti-inflammatory in the vasculature. To further define the cellular mechanisms necessary for vascular protection, we investigated how estrogen modulates hypoxic–adenosinergic vascular signaling responses and angiogenesis. Expression of estrogen receptors, purinergic mediators inclusive of adenosine, adenosine deaminase (ADA), and ATP were measured in human endothelial cells. Standard tube formation and wound healing assays were performed to assess angiogenesis in vitro. The impacts on purinergic responses in vivo were modeled using cardiac tissue from ovariectomized mice. CD39 and estrogen receptor alpha (ERα) levels were markedly increased in presence of estradiol (E2). Suppression of ERα resulted in decreased CD39 expression. Expression of ENT1 was decreased in an ER-dependent manner. Extracellular ATP and ADA activity levels decreased following E2 exposure while levels of adenosine increased. Phosphorylation of ERK1/2 increased following E2 treatment and was attenuated by blocking adenosine receptor (AR) and ER activity. Estradiol boosted angiogenesis, while inhibition of estrogen decreased tube formation in vitro. Expression of CD39 and phospho-ERK1/2 decreased in cardiac tissues from ovariectomized mice, whereas ENT1 expression increased with expected decreases in blood adenosine levels. Estradiol-induced upregulation of CD39 substantially increases adenosine availability, while augmenting vascular protective signaling responses. Control of CD39 by ERα follows on transcriptional regulation. These data suggest novel therapeutic avenues to explore in the amelioration of post-menopausal cardiovascular disease, by modulation of adenosinergic mechanisms.

Altered purinergic signaling in women undergoing cardiac surgery under cardiopulmonary bypass

“…[18,19] Our recent study also demonstrated the role of estrogen in purine expression in the setting of hypoxia, suggesting a more prominent role of purine dysfunction in our cardiac surgery patients. [20] The exaggerated in ammatory response in females, especially the levels of IL-6 and leptin, as compared to men, corroborates our nding of dysregulated ATP and adenosine. Level of leptin and IL6 have been shown to be elevated in heart failure and are well-studied markers of in ammation and oxidative stress.…”

“…[18,19] Some of our earlier work suggested a potential link between post-menopause and adenosine interaction in the setting of hypoxia, our work has shown that estrogen regulates CD39 increase under hypoxic conditions. [20] We hypothesize that purinergic signaling is dysregulated in older women resulting in adverse outcomes. We infer these pathways are disordered in the presence of decreased estrogen, which in the setting of the obesity and metabolic syndrome, exaggerate in ammation, increased brosis and diastolic dysfunction.…”