Hydrogen sulfide (H₂S) is a gasotransmitter that regulates both physiological and pathophysiological processes in mammalian cells. Recent studies have demonstrated that H₂S promotes aerobic energy production in the mitochondria in response to hypoxia, but its effect on anaerobic energy production has yet to be established. Glycolysis is the anaerobic process by which ATP is produced through the metabolism of glucose. Mammalian red blood cells (RBCs) extrude mitochondria and nucleus during erythropoiesis. These cells would serve as a unique model to observe the effect of H₂S on glycolysis-mediated energy production. The purpose of this study was to determine the effect of H₂S on glycolysis-mediated energy production in mitochondria-free mouse RBCs. Western blot analysis showed that the only H₂S-generating enzyme expressed in mouse RBCs is 3-mercaptopyruvate sulfurtransferase (MST). Supplement of the substrate for MST stimulated, but the inhibition of the same suppressed, the endogenous production of H₂S. Both exogenously administered H₂S salt and MST-derived endogenous H₂S stimulated glycolysis-mediated ATP production. The effect of NaHS on ATP levels was not affected by oxygenation status. On the contrary, hypoxia increased intracellular H₂S levels and MST activity in mouse RBCs. The mitochondria-targeted H₂S donor, AP39, did not affect ATP levels of mouse RBCs. NaHS at low concentrations (3–100 μM) increased ATP levels and decreased cell viability after 3 days of incubation in vitro. Higher NaHS concentrations (300–1000 μM) lowered ATP levels, but prolonged cell viability. H₂S may offer a cytoprotective effect in mammalian RBCs to maintain oxygen-independent energy production.

中文翻译：

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硫化氢对小鼠红细胞糖酵解能量产生的影响

硫化氢 (H ₂ S) 是一种气体递质，可调节哺乳动物细胞的生理和病理生理过程。最近的研究表明，H ₂ S 促进线粒体响应缺氧产生有氧能量，但其对无氧能量产生的影响尚未确定。糖酵解是通过葡萄糖代谢产生ATP的厌氧过程。哺乳动物红细胞 (RBC) 在红细胞生成过程中会挤出线粒体和细胞核。这些细胞将作为一个独特的模型来观察 H ₂ S 对糖酵解介导的能量产生的影响。本研究的目的是确定 H _2的影响S 对无线粒体小鼠红细胞中糖酵解介导的能量产生的影响。蛋白质印迹分析表明，在小鼠红细胞中表达的唯一 H ₂ S 生成酶是 3-巯基丙酮酸硫转移酶 (MST)。补充 MST 底物刺激，但抑制同样抑制 H 2 S 的内源性产生。外源性施用的_H 2 _S盐和 MST 衍生的内源性 H ₂ S 刺激糖酵解介导的 ATP 产生。NaHS 对 ATP 水平的影响不受氧合状态的影响。相反，缺氧增加了小鼠红细胞中的细胞内H _{2 S 水平和MST 活性。}线粒体靶向 H ₂S 供体 AP39 不影响小鼠红细胞的 ATP 水平。在体外培养 3 天后，低浓度 (3–100 μM) 的 NaHS 增加了 ATP 水平并降低了细胞活力。较高的 NaHS 浓度 (300–1000 μM) 降低了 ATP 水平，但延长了细胞活力。H ₂ S 可能在哺乳动物红细胞中提供细胞保护作用，以维持不依赖氧的能量产生。