Nonselective inhibition of neutrophil functions by sphinganine.
Nonselective inhibition of neutrophil functions by sphinganine.
复制标题
二氢鞘氨醇对中性粒细胞功能的非选择性抑制。
DOI：
10.1016/s0021-9258(18)61077-4
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
D. Lew
中科院分区：
文献类型：
--
作者：
D. Pittet;K. Krause;C. Wollheim;R. Bruzzone;D. Lew
鞘氨醇被认为是蛋白激酶C的特异性抑制剂。在本研究中，我们已经评估了鞘氨醇是否是一个方便的工具来探测蛋白激酶C在中性粒细胞功能中的作用。将人中性粒细胞加载荧光探针quin 2，然后平行测试细胞溶质游离Ca 2+、[Ca 2 +]i、膜电位变化、O2-产生和初级颗粒（含有β-葡萄糖醛酸酶）对各种刺激的胞吐作用。除了以剂量依赖性方式抑制O2-产生和胞吐作用外，鞘氨醇还阻断甲酰-甲硫氨酰-亮氨酰-苯丙氨酸诱导的[Ca 2 +]i瞬变。此外，鞘氨醇抑制胞吐引起的钙离子载体离子霉素。尽管二氢鞘氨醇由于佛波醇12-肉豆蔻酸酯13-乙酸酯而阻断了O2-的产生，但最引人注目的发现是该药物使细胞渗漏。因此，在与抑制细胞功能的浓度相似的浓度下，鞘氨醇显示导致细胞透化，如通过quin 2和细胞质标记物释放到细胞外介质中以及质膜电位的变化所评估的。因此，我们的结论是，鞘氨醇似乎不是一个合适的化合物的参与蛋白激酶C在中性粒细胞活化的评价。
Sphinganine has been proposed to be a specific inhibitor of protein kinase C. In the present study we have evaluated whether sphinganine is a convenient tool to probe for the role of protein kinase C in neutrophil function. Human neutrophils were loaded with the fluorescent probe quin2 and then tested in parallel for cytosolic free Ca2+, [Ca2+]i, membrane potential changes, O2- production, and exocytosis of primary granules (containing beta-glucuronidase) in response to various stimuli. In addition to inhibiting O2- production and exocytosis in a dose-dependent manner, sphinganine also blocked formyl-methionyl-leucyl-phenylalanine-induced [Ca2+]i, transients. Furthermore, sphinganine inhibited exocytosis elicited by the calcium ionophore ionomycin. Although sphinganine blocked O2- production due to phorbol 12-myristate 13-acetate, the most striking finding was that the drug rendered the cells leaky. Thus, at similar concentrations as those inhibiting cellular functions, sphinganine was shown to lead to cell permeabilization, as assessed by release of quin2 and cytoplasmic markers into the extracellular medium, and changes in plasma membrane potential. We conclude, therefore, that sphinganine does not appear to be a suitable compound for the evaluation of the involvement of protein kinase C in neutrophil activation.