Intravenous anesthetic propofol binds to 5-lipoxygenase and attenuates leukotriene B4 production
Intravenous anesthetic propofol binds to 5-lipoxygenase and attenuates leukotriene B4 production
复制标题
静脉麻醉异丙酚与 5-脂氧合酶结合并减弱白三烯 B4 的产生
作者：
Okuno;T.;Koutsogiannaki;S.;Ohba;M.;Chamberlain;M.;Bu;W.;Lin;F. Y.;Eckenhoff;R. G.;Yokomizo;T.,Yuki;K.
丙泊酚是一种静脉麻醉剂，主要通过CNS中的GABAA受体产生其麻醉作用，并且还减少N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）诱导的中性粒细胞呼吸爆发。由于fMLP刺激的中性粒细胞产生白三烯（LT）B4，我们研究了丙泊酚对体内和体外LTB 4产生的影响。在小鼠中进行盲肠结扎和穿刺手术，有或没有暴露于丙泊酚。异丙酚减弱了腹腔液中5-脂氧合酶（5-LOX）相关的花生四烯酸（AA）衍生物的产生。此外，在fMLP刺激的中性粒细胞和5-LOX转染的人胚肾细胞的体外实验中，丙泊酚减弱了5-LOX相关AA衍生物的产生。基于这些结果，我们假设丙泊酚会直接影响5-LOX功能。利用间位-阿齐-丙泊酚（AziPm）光标记了稳定的5-LOX蛋白，并研究了丙泊酚与5-LOX的结合位点。在5-LOX活性位点附近发现了两个丙泊酚结合口袋。丙氨酸扫描诱变进行5-LOX的残基在丙泊酚附近，我们评估这些口袋中的LTB 4生产的功能作用。我们证明，这些口袋是5-LOX活性的功能重要。这两个口袋可用于在未来探索新型5-LOX抑制剂。Okuno，T.，Koutsogiannaki，S.，Ohba，M.，张伯伦，M.，Bu，W.，Lin，F.-是的，埃肯霍夫河G.，横沟T.，尤基K.静脉麻醉剂丙泊酚与5-脂氧合酶结合并减弱白三烯B4的产生。
Propofol is an intravenous anesthetic that produces its anesthetic effect, largely via the GABAA receptor in the CNS, and also reduces the N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced neutrophil respiratory burst. Because fMLP-stimulated neutrophils produce leukotriene (LT)B4, we examined the effect of propofol on LTB4 production in vivo and in vitro. Cecal ligation and puncture surgery was performed in mice, with or without exposure to propofol. Propofol attenuated the production of 5-lipoxygenase (5-LOX)-related arachidonic acid (AA) derivatives in the peritoneal fluid. Also, in the in vitro experiments on fMLP-stimulated neutrophils and 5-LOX-transfected human embryonic kidney cells, propofol attenuated the production of 5-LOX-related AA derivatives. Based on these results, we hypothesized that propofol would directly affect 5-LOX function. Using meta-azi-propofol (AziPm), we photolabeled stable 5-LOX protein, which had been used to solve the X-ray crystallographic structure of 5-LOX, and examined the binding site(s) of propofol on 5-LOX. Two propofol binding pockets were identified near the active site of 5-LOX. Alanine scanning mutagenesis was performed for the residues of 5-LOX in the vicinity of propofol, and we evaluated the functional role of these pockets in LTB4 production. We demonstrated that these pockets were functionally important for 5-LOX activity. These two pockets can be used to explore a novel 5-LOX inhibitor in the future.—Okuno, T., Koutsogiannaki, S., Ohba, M., Chamberlain, M., Bu, W., Lin, F.-Y., Eckenhoff, R. G., Yokomizo T., Yuki, K. Intravenous anesthetic propofol binds to 5-lipoxygenase and attenuates leukotriene B4 production.