α-氰基-3,4-二羟基硫代肉桂酰胺对人中性粒细胞反应的抑制作用；一种蛋白酪氨酸激酶抑制剂。

Inhibition of human neutrophil responses by alpha-cyano-3,4-dihydroxythiocinnamamide; a protein-tyrosine kinase inhibitor.

作者信息

Dryden P, Duronio V, Martin L, Hudson A T, Salari H

机构信息

Department of Medicine, University of British Columbia, Vancouver, Canada.

出版信息

Br J Pharmacol. 1992 Jul;106(3):656-64. doi: 10.1111/j.1476-5381.1992.tb14391.x.

DOI:10.1111/j.1476-5381.1992.tb14391.x

PMID:1504749

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1907570/

Abstract

Activation of neutrophils results in increased tyrosine phosphorylation of several proteins that may have important roles in receptor/effector coupling. In this study, the effect of a protein tyrosine kinase inhibitor on receptor-mediated neutrophil activation by platelet-activating factor (PAF), leukotriene, B4 (LTB4) and N-formylmethionylleucylphenylalanine (FMLP) is investigated. 2. alpha-Cyano-3,4-dihydroxythiocinnamamide dose-dependently inhibited intracellular calcium release and superoxide generation from human neutrophils activated by 1 microM LTB4, PAF, and FMLP. 3. In the presence of cytochalasin B, FMLP stimulated elastase release from neutrophils was also inhibited to unstimulated levels by 5 min pretreatment with alpha-cyano-3,4-dihydroxythiocinnamamide. 4. The inhibitory action of alpha-cyano-3,4-dihydroxythiocinnamamide was found to be at or upstream of phospholipase C activation, blocking both phosphatidylinositol hydrolysis and protein kinase C activation. alpha-Cyano-3,4-dihydroxythiocinnamamide did not affect agonist receptor binding sites or receptor affinity in neutrophils. 5. Immunoblot analysis demonstrated the tyrosine phosphorylation of proteins of 41, 56, 66, and 104 kDa in neutrophils treated with agonists. Treatment of neutrophils with alpha-cyano-3,4-dihydroxythiocinnamamide prior to stimulation with chemoattractants reduced tyrosine phosphorylation of the above phosphoproteins. 6. These results indicate that alpha-cyano-3,4-dihydroxythiocinnamamide might be a useful agent in characterizing the essential proteins and biochemical pathways that regulate neutrophil activation.

摘要

中性粒细胞的激活导致几种蛋白质的酪氨酸磷酸化增加，这些蛋白质可能在受体/效应器偶联中发挥重要作用。在本研究中，研究了一种蛋白质酪氨酸激酶抑制剂对血小板活化因子（PAF）、白三烯B4（LTB4）和N-甲酰甲硫氨酰亮氨酰苯丙氨酸（FMLP）介导的中性粒细胞激活的影响。2. α-氰基-3,4-二羟基硫代肉桂酰胺剂量依赖性地抑制了由1μM LTB4、PAF和FMLP激活的人中性粒细胞的细胞内钙释放和超氧化物生成。3. 在细胞松弛素B存在的情况下，用α-氰基-3,4-二羟基硫代肉桂酰胺预处理5分钟，FMLP刺激的中性粒细胞弹性蛋白酶释放也被抑制到未刺激水平。4. 发现α-氰基-3,4-二羟基硫代肉桂酰胺的抑制作用在磷脂酶C激活处或上游，阻断磷脂酰肌醇水解和蛋白激酶C激活。α-氰基-3,4-二羟基硫代肉桂酰胺不影响中性粒细胞中激动剂受体结合位点或受体亲和力。5. 免疫印迹分析表明，用激动剂处理的中性粒细胞中41、56、66和104 kDa蛋白质发生酪氨酸磷酸化。在用趋化因子刺激之前，用α-氰基-3,4-二羟基硫代肉桂酰胺处理中性粒细胞可降低上述磷蛋白的酪氨酸磷酸化。6. 这些结果表明，α-氰基-3,4-二羟基硫代肉桂酰胺可能是一种有用的试剂，可用于表征调节中性粒细胞激活的必需蛋白质和生化途径。