P2Y11 Purinoceptor Mediates the ATP-Enhanced Chemotactic Response of Rat Neutrophils
P2Y11 Purinoceptor Mediates the ATP-Enhanced Chemotactic Response of Rat Neutrophils
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DOI：
10.1254/jphs.12173fp
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发表时间：
2012-12-01
影响因子：
3.5
通讯作者：
Azuma, Yukio
中科院分区：
文献类型：
--
作者：
Alkayed, Feras;Kashimata, Masanori;Azuma, Yukio
ATP及其水解产物如腺苷通过激活嘌呤受体和腺苷受体调节中性粒细胞的趋化性。本研究旨在检测外源性ATP、嘌呤受体激活和A（3）腺苷受体激活作为控制大鼠中性粒细胞定向和迁移的信号级联的关键步骤。这些步骤中的一个或多个可能是治疗炎性疾病的潜在治疗靶点。用N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）刺激大鼠中性粒细胞的趋化性，并用EZ-TAXIScan装置测定。研究了腺苷三磷酸双磷酸酶、外源性ATP、苏拉明（P2 X和P2 Y阻断剂）、PPADS（P2 X阻断剂）、TNP-ATP（P2 X（1）和P2 X（3）拮抗剂）和活性蓝2（P2 Y阻断剂）对趋化反应的影响。大鼠中性粒细胞趋化性显着抑制腺苷三磷酸双磷酸酶。fMLP诱导的大鼠中性粒细胞趋化性被ATP增强，被苏拉明阻断，不受PPADS或TNP-ATP的影响，并且被RB-2显著抑制。Western blotting结果显示A（3）、P2 Y（2）和P2 Y（11）在大鼠中性粒细胞中表达。ATP通过P2 Y（11）嘌呤受体而不是P2 X嘌呤受体或A（3）腺苷受体增强大鼠中性粒细胞对fMLP刺激的趋化反应，并且该反应在宿主防御和致病性中起关键作用。[补充电影：仅在http：//dx.doi.org/10.1254/jphs.12173FP上提供]
ATP and hydrolysis products of ATP like adenosine regulate the chemotaxis of neutrophils by activating purinoceptors and adenosine receptors. The present study was designed to examine exogenous ATP, activation of purinoceptors, and activation of A(3) adenosine receptor as key steps in the signal cascades that control cell orientation and migration of rat neutrophils. One or more of those steps might be potential therapeutic targets for treatment of inflammatory diseases. The chemotaxis of rat neutrophils was stimulated with N-formyl-methionyl-leucyl-phenylalanine (fMLP) and measured with an EZ-TAXIScan apparatus. The effects of apyrase, exogenous ATP, suramin (P2X and P2Y blocker), PPADS (a P2X blocker), TNP-ATP (P2X(1) and P2X(3) antagonist), and Reactive Blue 2 (a P2Y blocker) on the chemotactic response were also investigated. Rat neutrophil chemotaxis was significantly suppressed by apyrase. fMLP induced rat neutrophil chemotaxis was potentiated by ATP, blocked by suramin, not affected by PPADS or TNP-ATP, and significantly inhibited by RB-2. Western blotting showed that A(3), P2Y(2), and P2Y(11) were expressed in rat neutrophils. The chemotactic response of rat neutrophils to fMLP stimulation is potentiated by ATP via P2Y(11) purinoceptors but not P2X purinoceptors or A(3) adenosine receptor, and that the response plays a critical role in host defense and pathogenicity. [Supplementary Movies: available only at http://dx.doi.org/10.1254/jphs.12173FP]