Abstract—Metabolic stress caused by a lack of glucose significantly affects the state of red blood cells, where glycolysis is the main pathway for the production of ATP. Hypoglycemia can be both physiological (occurring during fasting and heavy physical exertion) and pathological (accompanying a number of diseases, such as diabetes mellitus). In this study, we have characterized the state of isolated erythrocytes under metabolic stress caused by the absence of glucose. It was established that 24 h of incubation of the erythrocytes in a glucose-free medium to simulate blood plasma led to a two-fold decrease in the ATP level into them. The cell size, as well as intracellular sodium concentration increased. These findings could be the result of a disruption in ion transporter functioning because of a decrease in the ATP level. The calcium level remained unchanged. With a lack of glucose in the medium of isolated erythrocytes, there was no increase in ROS and a significant change in the level of nitric oxide, while the level of the main low-molecular weight thiol of cells, glutathione (GSH) decreased by almost 2 times. It was found that the metabolic stress of isolated red blood cells induced hemoglobin glutathionylation despite the absence of ROS growth. The cause was the lack of ATP, which led to a decrease in the level of GSH because of the inhibition of its synthesis and, probably, due to a decrease in the NADPH level required for glutathione (GSSG) reduction and protein deglutathionylation. Thus, erythrocyte metabolic stress induced hemoglobin glutathionylation, which is not associated with an increase in ROS. This may have an important physiological significance, since glutathionylation of hemoglobin changes its affinity for oxygen.

摘要：葡萄糖缺乏引起的代谢应激显着影响红细胞的状态，其中糖酵解是产生 ATP 的主要途径。低血糖可以是生理性的（在禁食和剧烈体力消耗时发生），也可以是病理性的（伴随许多疾病，例如糖尿病）。在这项研究中，我们描述了在缺乏葡萄糖引起的代谢应激下分离的红细胞的状态。已经确定，将红细胞在无葡萄糖培养基中培养 24 小时以模拟血浆，会导致红细胞中 ATP 水平下降两倍。细胞大小以及细胞内钠浓度增加。这些发现可能是由于 ATP 水平下降而导致离子转运蛋白功能中断的结果。钙水平保持不变。分离红细胞培养基中缺乏葡萄糖时，ROS没有增加，一氧化氮水平也没有显着变化，而细胞的主要低分子量硫醇谷胱甘肽（GSH）水平几乎下降了2次。研究发现，尽管没有ROS生长，分离的红细胞的代谢应激仍会诱导血红蛋白谷胱甘肽化。原因是缺乏 ATP，导致 GSH 水平下降，因为其合成受到抑制，并且可能是由于谷胱甘肽 (GSSG) 还原和蛋白质去谷胱甘肽化所需的 NADPH 水平下降。因此，红细胞代谢应激诱导血红蛋白谷胱甘肽化，这与 ROS 的增加无关。这可能具有重要的生理意义，因为血红蛋白的谷胱甘肽化改变了其对氧的亲和力。