Acute and chronic suppression of leukotriene B4 synthesis ex vivo in neutrophils from patients with rheumatoid arthritis beginning treatment with methotrexate.
Acute and chronic suppression of leukotriene B4 synthesis ex vivo in neutrophils from patients with rheumatoid arthritis beginning treatment with methotrexate.
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开始接受甲氨蝶呤治疗的类风湿性关节炎患者的中性粒细胞离体白三烯 B4 合成受到急性和慢性抑制。
DOI：
10.1002/art.1780350403
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发表时间：
1992
影响因子：
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通讯作者：
Weinblatt,ME
中科院分区：
文献类型：
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作者：
Sperling,RI;Benincaso,AI;Anderson,RJ;Coblyn,JS;Austen,KF;Weinblatt,ME
目的：比较口服甲氨蝶呤（MTX）治疗的累积效应（6-8周后）（给药后24小时）通过5-脂氧合酶（5-LO）途径对来自开始用MTX治疗的活动性类风湿性关节炎（RA）患者的中性粒细胞中花生四烯酸代谢的影响。结果：在6 - 8周剂量前，用钙离子载体A23187体外激活的中性粒细胞中总免疫反应性白三烯B4（LTB 4）形成显著抑制（33%），与首次给药前水平相比（给药前6-8周平均值± SEM 8.29 ± 1.24 ng/106个细胞vs第1次给药前12.29 ± 2.13 ng/106个细胞;P= 0.03）。与相应的给药前水平相比，首次给药后（27%;P= 0.07）和6-8周给药后（43%;P= 0.05）也观察到降低。MTX处理显著减少了钙离子载体活化的中性粒细胞产生的5-LO途径产物（5-羟基二十碳四烯酸+ 6-反式-LTB 4 + LTB 4 + LTB 4的ω-氧化产物）的总量，通过反相高效液相色谱分离后的积分光密度进行定量。与首次给药前水平相比，首次给药后（26%;P= 0.025）、第6- 8周给药前即刻（23%;P= 0.05）和第6-8周给药后（47%;P= 0.0033）观察到降低，与第1次给药前水平相比，第6-8周给药后与给药前水平相比（32%;P= 0.04）观察到降低。MTXtherapy.Conclusion.LTB4的ω-氧化产物的形成和中性粒细胞5-LO途径产物的总生成量的显著减少，而3 H-花生四烯酸的释放或血小板活化因子的生成没有显著变化，表明中性粒细胞中5-LO酶的活性受到抑制。我们得出结论，活动性RA患者每周口服MTX治疗可抑制中性粒细胞5-LO途径产物的生成，其模式与抑制5-LO酶活性一致;首次给药后观察到效果。5-LO的抑制具有细胞选择性和累积性，每周MTX给药后观察到叠加的增量抑制。
Objective.To compare the cumulative effects of oral methotrexate (MTX) therapy (after 6–8 weeks) with the acute effects (24 hours after a dose) on arachidonic acid metabolism by the 5‐lipoxygenase (5‐LO) pathway in neutrophils from patients with active rheumatoid arthritis (RA) who were beginning therapy with MTX.Methods.Neutrophils and monocytes were isolated from whole blood from 7 patients with RA, immediately before and 24 hours after their first weekly dose of 7.5 mg of MTX, and again after their dose at 6–8 weeks.Results.Total immunoreactive leukotriene B4(LTB4) formation in neutrophils activated ex vivo with calcium ionophore A23187 was significantly suppressed (by 33%) before the 6–8‐week dose, compared with the level before the first dose (mean ± SEM 8.29 ± 1.24 ng/106cells at predose 6–8 weeks versus 12.29 ± 2.13 ng/106cells at predose 1;P= 0.03). Reductions were also observed after the first dose (27%;P= 0.07) and after the 6–8‐week dose (43%;P= 0.05) compared with the respective predose levels. MTX treatment produced significant reductions in the total generation of 5‐LO pathway products (5‐hydroxyeicosatetraenoic acid + 6‐trans‐LTB4+ LTB4+ ω‐oxidation products of LTB4) by calcium ionophore‐activated neutrophils, as quantitated by integrated optical density after resolution on reverse‐phase high‐performance liquid chromatography. Decreases were observed after the first dose (26%;P= 0.025), immediately before the 6–8‐week dose (23%;P= 0.05), and after the 6–8‐week dose (47%;P= 0.0033) compared with levels before the first dose, and after the 6–8‐week dose compared with the level before it (32%;P= 0.04). The generation of LTB4by calcium ionophore‐activated monocytes was not significantly affected by MTX therapy.Conclusion.The significant decreases in the formation of ω‐oxidation products of LTB4and in the total generation of neutrophil 5‐LO pathway products in the absence of a significant change in the release of3H‐arachidonic acid or the generation of platelet‐activating factor suggest that the activity of the 5‐LO enzyme in neutrophils is inhibited. We conclude that weekly oral MTX therapy in patients with active RA inhibits neutrophil 5‐LO pathway product generation in a pattern consistent with inhibition of the activity of the 5‐LO enzyme; an effect is observed after the first dose. The inhibition of 5‐LO is cell‐selective and cumulative, with a superimposed incremental inhibition observed after the weekly MTX dose.