Inhibition of Suicidal Erythrocyte Death by Chronic Hypoxia

High-altitude polycythemia is defined by the increase of hematocrit and hemoglobin at high altitudes caused by production of excessive erythrocytes. Eryptosis is a process by which mature erythrocytes undergo self-destruction sharing several features with apoptosis. However, the eryptosis in high-altitude hypobaric hypoxia is unknown. Thus, the main purpose of this study was to investigate whether chronic hypoxia affected eryptosis and, if so, by what mechanisms. Biotin labeling technology was utilized to study the survival of red blood cells in chronic hypoxia. Flow cytometry was used to determine the volume of mature erythrocytes from forward scatter, phosphatidylserine scrambling from annexin-V-binding, intracellular Ca²⁺ from Fluo-3-AM, reactive oxygen species (ROS) abundance from ROS-probe, and ceramide and CD47 abundance utilizing specific antibodies. The volume of mature erythrocytes was significantly changed, and the percentage of annexin-V-binding cells was significantly decreased under chronic hypobaric hypoxia. Erythrocyte survival was improved under chronic hypoxia, and chronic hypoxia resulted in a decrease in intracellular Ca²⁺in vivo and influenced eryptosis which was induced by the Ca²⁺-ionophore ionomycin (1 μM, 60 minutes) in vitro. Chronic hypoxia also resulted in an increase in CD47 and ceramide abundance, but it had no effect on ROS formation. Chronic hypobaric hypoxia can inhibit eryptosis by decreasing intracellular Ca²⁺ and increasing integrin-associated protein CD47.