2025 – Physiological oxygen levels reset K+ channel activity in human vascular endothelial cells

Human endothelial cells (EC) play a critical role in vascular homeostasis and their function is influenced by oxygen tension. This study investigates for the first time the effects of long-term adaptation (5 days) of two major EC types to physiological oxygen tension (5kPa) on basal and nitric oxide (NO)-modulated K⁺ channel activities. Whole-cell patch clamp experiments demonstrate that human umbilical vein EC (HUVEC) exhibit larger basal K⁺ outward and smaller inward currents under 5kPa O₂ compared to standard hyperoxic (18kPa) culture conditions. Outward currents were potentiated by NO only under hyperoxia. Human cerebral microvascular EC (hCMEC/D3) showed larger outward currents under 5kPa O₂ which were further potentiated by NO. Pharmacological isolation of different K⁺ currents using tetraethylammonium, TRAM-34 and apamin revealed differential effects in EC adapted to 5kPa or 18kPa O₂. Under 5kPa O₂, both cell types show greater contributions of TEA-sensitive currents and in addition hCMEC/D3 cells exhibit higher proportions of TRAM-34 and apamin-sensitive currents under 5kPa O₂. In HUVEC, changes in half-activation voltage and hyperpolarized membrane potentials were detected only under hyperoxic conditions following NO exposure, with both cell types exhibiting altered current activation kinetics of outward and inward currents. Notably, expression of KCa3.1, KCa1.1, KCa2.3 and Kir6.1 channels was unaffected by O₂, suggesting that changes in whole-cell currents in both EC types were due to channel modulation. Thus, our findings reveal that physiological O₂ tension shapes the electrophysiological phenotype of human EC by modulating K⁺ channel function and NO responsiveness. The novel insights into the modulation of EC K⁺ channels by O₂ has implications for the regulation of vascular tone and design and use of experimental models in vitro for high throughput drug discovery and clinical translation.