Spragg R G, Hinshaw D B, Hyslop P A, Schraufstätter I U, Cochrane C G
J Clin Invest. 1985 Oct;76(4):1471-6. doi: 10.1172/JCI112126.
To investigate mechanisms whereby oxidant injury of cells results in cell dysfunction and death, cultured endothelial cells or P388D1 murine macrophage-like 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 less than 30% of control values within 3 min of exposure to 5 mM 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.
为了研究细胞的氧化损伤导致细胞功能障碍和死亡的机制，将培养的内皮细胞或P388D1鼠巨噬细胞样细胞暴露于包括H2O2、O2-（由黄嘌呤的酶促氧化产生）在内的氧化剂中，或暴露于受刺激的多形核白细胞（PMN）中。尽管在30分钟前台盼蓝排斥率没有降低，但发现暴露于5 mM H2O2后3分钟内细胞ATP降至对照值的30%以下。受刺激的PMN加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增加了170%，AMP增加了900%，ATP损失83%的同时，肌苷一磷酸（IMP）和肌苷化学计量增加。计算得出的能荷[（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）导致ATP下降速率与H2O2诱导的相似。此外，暴露于寡霉素加2-脱氧葡萄糖诱导的核苷酸变化在性质上与氧化剂诱导的相似。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ATP消耗增加来解释。这些结果表明，H2O2导致细胞ATP水平迅速而显著下降，类似于代谢抑制剂阻止ATP产生时所观察到的情况。
