ALTERATIONS IN ADENOSINE-TRIPHOSPHATE AND ENERGY-CHARGE IN CULTURED ENDOTHELIAL AND P388D1 CELLS AFTER OXIDANT INJURY
ALTERATIONS IN ADENOSINE-TRIPHOSPHATE AND ENERGY-CHARGE IN CULTURED ENDOTHELIAL AND P388D1 CELLS AFTER OXIDANT INJURY
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DOI：
10.1172/jci112126
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
COCHRANE, CG
中科院分区：
文献类型：
--
作者：
SPRAGG, RG;HINSHAW, DB;COCHRANE, CG
为了研究细胞氧化损伤导致细胞功能障碍和死亡的机制，将培养的 P388D1 小鼠巨噬细胞样细胞内皮细胞暴露于氧化剂，包括 H2O2、O2-（由黄嘌呤酶氧化产生），或暴露于刺激的多形核白细胞 (PMN)。尽管台盼蓝排除在 30 分钟前并未减弱，但发现细胞 ATP 在暴露于 5 mH H2O2 的 3 分钟内降至对照值的 < 30%。受刺激的 PMN 加上 P388D1 导致细胞 ATP 水平下降 50%。在氧化损伤的最初几分钟内，细胞的总腺苷酸含量下降了 85%。细胞 ADP 增加 170%，AMP 增加 900%，ATP 损失 83%，同时 IMP 和肌苷化学计量增加。计算出的能量消耗 [(ATP + 1/2 AMP)/(ATP + ADP + AMP)] 从 0.95 降至 0.66。将 P388D1 暴露于寡霉素加 2-脱氧葡萄糖（分别抑制 ATP 的氧化和糖酵解生成）会导致 ATP 下降速率与 H2O2 诱导的速率相似。此外，暴露于寡霉素加2-脱氧葡萄糖诱导的核苷酸改变在质量上与氧化剂诱导的改变相似。细胞腺苷酸的损失不能用嘌呤从头合成的停滞或 Na+-K+ ATP 酶或线粒体 F0-ATP 酶导致的 ATP 消耗增加来解释。这些结果表明，H2O2 会导致细胞 ATP 水平快速而大幅下降，类似于代谢抑制剂抑制 ATP 产生时所观察到的情况。
To investigate mechanisms whereby oxidant injury of cells results in cell dysfunction and death, cultured endothelial cells of P388D1 murine macrophagelike cells were exposed to oxidants including H2O2, O2- (generated by the enzymatic oxidation of xanthine), or to stimulated polymorphonuclear leukocytes (PMN). Although Trypan Blue exclusion was not diminished before 30 min, cellular ATP was found to fall to < 30% of control values within 3 min of exposure to 5 mH H2O2. Stimulated PMN plus P388D1 caused a 50% fall in cellular ATP levels. During the first minutes of oxidant injury, total adenylate content of cells fell by 85%. Cellular ADP increased 170%, AMP increased 900%, and an 83% loss of ATP was accompanied by a stoichiometric increase in IMP and inosine. Calculated energy charge [(ATP + 1/2 AMP)/(ATP + ADP + AMP)] fell from 0.95 to 0.66. Exposure of P388D1 to oligomycin plus 2-deoxyglucose (which inhibit oxidative and glycolytic generation of ATP, respectively) resulted in a rate of ATP fall similar to that induced by H2O2. In addition, nucleotide alterations induced by exposure to oligomycin plus 2-deoxyglucose were qualitatively similar to those induced by the oxidant. Loss of cell adenylates could not be explained by arrest of de novo purine synthesis or increased ATP consumption by the Na+-K+ ATPase or the mitochondrial F0-ATPase. These results indicate that H2O2 causes a rapid and profound fall in cellular ATP levels similar to that seen when ATP production is arrested by metabolic inhibitors.