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Hypothermic injury: the mitochondrial calcium, ATP and ROS love-hate triangle out of balance

机译:低温伤害:线粒体钙,ATP和ROS的爱恨三角失去平衡

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Background/Aims: Catecholamines prevent hypothermic cell death which accounts for severe tissue damage and impaired allograft function after prolonged organ preservation. Here, we identified cellular processes which govern hypothermia-mediated cell death in endothelial cells and how they are influenced by dopamine. Methods: Lactate dehydrogenase assay, intracellular ATP, reactive oxygen species and reduced thio-group measurement, intracellular calcium measurement and mitochondrial calcium staining were performed in the study. Results: Intracellular ATP was almost completely depleted within 12 hrs of hypothermic preservation in untreated human umbilical vein endothelial cells (HUVEC), while dopamine pre-treatment significantly delayed ATP depletion. 4 hrs after hypothermia a redox imbalance was observed in untreated cells, which increased with the duration of hypothermia. The redox imbalance was primarily caused by depletion of SH reduction equivalents and was significantly inhibited by dopamine. In addition, hypothermia-induced Ca2+ influx and mitochondrial Ca2+ accumulation were both prevented by dopamine. The protective effect of dopamine was abrogated by ionomycin and sodium azide and partly by oligomycin and CCCP. Conclusions: Our data demonstrated that loss of intracellular ATP, generation of a redox imbalance and accumulation of intracellular Ca2+ underlie cold preservation injury. Dopamine improves the redox balance, prevents intracellular Ca2+ accumulation and delays ATP depletion.
机译:背景/目的:儿茶酚胺可防止低温细胞死亡,这是造成器官长期保存后严重组织损伤和同种异体移植功能受损的原因。在这里,我们确定了控制低温介导的内皮细胞死亡的细胞过程以及它们如何受到多巴胺的影响。方法:进行乳酸脱氢酶测定,细胞内ATP,活性氧和还原硫基测定,细胞内钙测定和线粒体钙染色。结果:未经处理的人脐静脉内皮细胞(HUVEC)在低温保存后12小时内,细胞内ATP几乎完全耗尽,而多巴胺预处理显着延迟了ATP耗尽。体温过低4小时后,未处理的细胞中出现了氧化还原失衡,这种现象随着体温持续时间的增加而增加。氧化还原失衡主要是由于减少SH还原当量引起的,并且被多巴胺显着抑制。此外,多巴胺可防止体温过低引起的Ca2 +内流和线粒体Ca2 +的积累。多巴胺的保护作用被离子霉素和叠氮化钠取消,部分被寡霉素和CCCP取消。结论:我们的数据表明,细胞内ATP的损失,氧化还原失衡的产生和细胞内Ca2 +的积累是冷藏损害的基础。多巴胺可改善氧化还原平衡,防止细胞内Ca2 +积累并延迟ATP消耗。

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