首页> 外文期刊>Endothelium: Journal of endothelial cell research >pH-dependent oxidant production following inhibition of the mitochondrial electron transport chain in pulmonary endothelial cells.
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pH-dependent oxidant production following inhibition of the mitochondrial electron transport chain in pulmonary endothelial cells.

机译:在肺内皮细胞中抑制线粒体电子传输链后,pH依赖的氧化剂产生。

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We investigated the effect of changes in intracellular pH (pHi) and Na/H antiport activity on intracellular oxidant production in human pulmonary artery endothelial cells (HPAEC) following disruption of cellular metabolism. Oxidant production was measured with oxidant-sensitive probes (2',7'-dichlorofluorescein diacetate [H2DCF], dihydroethidium [DHE]) following treatment with inhibitors of mitochondrial electron transport and glycolysis (antimycin/2-deoxyglucose, A/D). A/D treatment increased oxidant production in a dose-dependent fashion over 2 hours. Omission of 2-deoxyglucose did not alter the magnitude of oxidant production. Inhibition at more proximal sites in the mitochondrial electron transport chain inhibited oxidant production. These data suggested that the mitochondrial electron transport chain was the source of oxidant production. Fluorescent imaging experiments confirmed the mitochondrial origin of the increased oxidant production under these conditions. Maneuvers that reduced pHi and inhibited Na/H exchange (acidosis, specific Na/H exchange inhibitors) attenuated oxidant production, whereas maneuvers that raised pHi (monensin) potentiated oxidant production. The results with the pH-insensitive probe (DHE) confirmed that oxidant production was pH-dependent. Oxidant production preceded significant loss of cell viability at 6 h following A/D treatment. These results demonstrate that oxidant production following inhibition of mitochondrial electron transport in HPAEC is pH-dependent and may contribute to endothelial cell injury by increasing endogenous oxidative stress.
机译:我们调查了细胞内pH(pHi)和Na / H反端口活性变化对细胞代谢中断后人肺动脉内皮细胞(HPAEC)细胞内氧化剂产生的影响。在用线粒体电子转运和糖酵解抑制剂(抗霉素/ 2-脱氧葡萄糖,A / D)处理后,用氧化剂敏感的探针(2',7'-二氯荧光素二乙酸盐[H2DCF],二氢乙啶[DHE])测量氧化剂的产生。 A / D处理在2小时内以剂量依赖的方式增加了氧化剂的产生。省略2-脱氧葡萄糖不会改变氧化剂的产生量。线粒体电子传输链中更近端的抑制作用抑制了氧化剂的产生。这些数据表明线粒体电子传输链是氧化剂产生的来源。荧光成像实验证实了在这些条件下增加的氧化剂产生的线粒体起源。降低pHi并抑制Na / H交换(酸中毒,特定的Na / H交换抑制剂)的操作会减弱氧化剂的产生,而提高pHi(monensin)的操作会增强氧化剂的产生。 pH不敏感探针(DHE)的结果证实了氧化剂的产生是pH依赖性的。 A / D处理后6小时,氧化剂的产生先于细胞活力的明显丧失。这些结果证明,在HPAEC中抑制线粒体电子运输后的氧化剂产生是pH依赖性的,并且可能通过增加内源性氧化应激而导致内皮细胞损伤。

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