Activation of kinin B1 receptors induces chemotaxis of human neutrophils
Activation of kinin B1 receptors induces chemotaxis of human neutrophils
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DOI：
10.1189/jlb.1205744
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发表时间：
2006-07-01
影响因子：
5.5
通讯作者：
Figueroa, C. D.
中科院分区：
文献类型：
--
作者：
Ehrenfeld, P.;Millan, C.;Figueroa, C. D.
激肽是生物活性肽，是细胞炎症的强有力介质。它们通过诱导血管舒张和增加血管通透性和疼痛来模拟炎症的主要体征。中性粒细胞通过几种刺激被化学吸引到炎症部位。然而，关于激肽肽的趋化作用的证据一直是矛盾的。我们分析了激肽131受体激动剂对健康人外周血中性粒细胞的趋化作用。在琼脂糖技术下使用迁移进行趋化。为了测试B受体激动剂的作用，在37 ℃下在5%CO2 -95%空气中对用1 ng/ml白细胞介素-1 β致敏的中性粒细胞进行每种测定过夜。使用未引发的细胞或用forinyl-Met-Leu-Phe（fMLP）攻击的细胞进行同时实验。当用10（-10）M的Lys-des[Arg（9）]-缓激肽（LDBK）或des[Arg（9）]-缓激肽攻击致敏的中性粒细胞时，观察到明显的趋化活性，但当使用未致敏的细胞时则没有。在0.5 mM放线菌酮和10 μ g/ml布雷菲德菌素A存在下引发细胞后，观察到趋化反应减少，表明需要某些蛋白质生物合成。逆转录-聚合酶链反应和原位杂交等技术证实了B受体mRNA的表达，免疫细胞化学和放射自显影也证实了这一点。B-1受体蛋白的表达。与其他化学引诱剂如fMLP相反，细胞溶质细胞内钙对B-1受体激动剂LDBK的反应不增加。在急性炎症的早期阶段产生的激肽B受体激动剂可能有利于中性粒细胞向炎症部位的募集。
Kinins are biologically active peptides that are powerful mediators of cellular inflammation. They mimic the cardinal signs of inflammation by inducing vasodilatation and by increasing vascular permeability and pain. Neutrophils are chemoattracted to sites of inflammation by several stimuli. However, the evidence concerning the chertiotactic effect of kinin peptides has been contradictory. We analyzed the chemotactic effect of kinin 131 receptor agonists on nentrophils isolated from peripheral blood of human healthy subjects. Chemotaxis was performed using the migration under agarose technique. To test the effect of B, receptor agonists, each assay was carried out overnight at 37 degrees C in 5% CO2-95% air on nentrophils primed with I ng/ml interleukin-1 beta. Simultaneous experiments were performed using unprimed cells or cells challenged with forinyl-Met-Leu-Phe (fMLP). A clear chemotactic activity was observed when primed nentrophils were challenged with Lys-des[Arg(9)]-bradykinin (LDBK) or des[Arg(9)]-bradykinin at 10(-10) M but not when unprimed cells were used. A reduction in the chemotactic response was observed after priming of cells in the presence of 0.5 mM cycloheximide and 10 mu g/ml brefeldin A, suggesting that some protein biosynthesis is required. Techniques such as reverse transcriptase-polymerase chain reaction and in situ hybridization confirmed the expression of the B, receptor mRNA, and immunocytochemistry and autoradiography demonstrated. the expression of the B-1 receptor protein. In contrast to other chemoattractants such as fMLP, cytosolic intracellular calcium did not increase in response to die B-1 receptor agonist LDBK. A generation of kinin B, receptor agonists during the early phase of acute inflammation may favor the recruitment of neutrophils to the inflammatory site.