MODULATION OF INTRACELLULAR FORMATION OF REACTIVE OXYGEN INTERMEDIATES IN PERITONEAL-MACROPHAGES AND MICROGLIA/BRAIN MACROPHAGES BY PROPENTOFYLLINE
MODULATION OF INTRACELLULAR FORMATION OF REACTIVE OXYGEN INTERMEDIATES IN PERITONEAL-MACROPHAGES AND MICROGLIA/BRAIN MACROPHAGES BY PROPENTOFYLLINE
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DOI：
10.1038/jcbfm.1994.19
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发表时间：
1994-01-01
影响因子：
6.3
通讯作者：
KREUTZBERG, GW
中科院分区：
文献类型：
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作者：
BANATI, RB;SCHUBERT, P;KREUTZBERG, GW
缺血诱导的神经细胞死亡可以部分预防丙戊茶碱，结构上与黄嘌呤衍生物，与内源性腺苷的神经调节功能相互作用的药理学。为了评价丙戊茶碱的神经保护作用的可能机制，我们研究了其对小胶质细胞中活性氧中间体的细胞产生的影响，与腹膜巨噬细胞相比，小胶质细胞在病理条件下可以分化为脑巨噬细胞。使用流式细胞仪测定，我们确定了细胞内形成的活性氧中间体，通过测量氧化的膜渗透性和nonfluorescent二氢罗丹明123的阳离子和胞内捕获，绿色荧光罗丹明123在单个活细胞。丙戊茶碱在50 μ M的治疗浓度完全抑制钙依赖性刀豆蛋白A诱导的活性氧中间体在腹腔巨噬细胞的生产增加。在分离和培养的小胶质细胞，其中有一个高的自发呼吸爆发活动，活性氧中间体的自发生产减少了约30%。阿佛波酯12-肉豆蔻酸酯13-乙酸酯诱导的呼吸爆发活性的上升不能抑制propentofylline在任何一种细胞类型。活性氧中间体的产生增加被认为有助于脑缺血、水肿和神经退行性疾病如阿尔茨海默病后的神经细胞死亡。这些病理条件都伴随着小胶质细胞的激活。因此，我们认为，propentofylline的神经保护特性可能部分是由于调制的小胶质细胞产生的潜在有害的活性氧中间体。
Ischemia-induced nerve cell death can partly be prevented by propentofylline, a pharmacon structurally related to xanthine derivates that interacts with the neuromodulatory function of endogenous adenosine. To evaluate a possible mechanism of neuroprotection by propentofylline, we studied its effect on the cellular production of reactive oxygen intermediates in microglial cells, which under pathological conditions can differentiate into brain macrophages, in comparison to peritoneal macrophages. Using a flow cytometric assay, we determined the intracellular formation of reactive oxygen intermediates by measuring the oxidation of the membrane-permeable and nonfluorescent dihydrorhodamine 123 to the cationic and intracellularly trapped, green fluorescent rhodamine 123 in single viable cells. Propentofylline at the therapeutic concentration of 50 mu M completely inhibited the Ca2+-dependent Con A-induced increase in the production of reactive oxygen intermediates in peritoneal macrophages. In isolated and cultured microglial cells, which have a high spontaneous respiratory burst activity, the spontaneous production of reactive oxygen intermediates was reduced by similar to 30%. A phorbol 12-myristate 13-acetate-induced rise in the respiratory burst activity could not be inhibited by propentofylline in either cell type. An increased generation of reactive oxygen intermediates is thought to contribute to nerve cell death after brain ischemia, edema, and neurodegenerative diseases like Alzheimer's disease. These pathological conditions are all accompanied by an activation of microglial cells. We therefore suggest that the neuroprotective properties of propentofylline might in part be due to a modulation of the microglial production of potentially harmful reactive oxygen intermediates.