Evidence of a role for cyclic ADP-ribose in calcium signaling and migration in human neutrophils

环 ADP-核糖在人中性粒细胞钙信号传导和迁移中作用的证据

基本信息

批准号：
15591967
负责人：
MORITA Katsuya
金额：
$ 1.92万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

Cyclic ADP-ribose (cADPR) has a powerful Ca^<2+>-mobilizing action and behaves as a second messenger of many agonists, thereby modulating a wide number of Ca^<2+>-mediated cell processes. CD38, the best-characterized mammalian ADP-ribosyl cyclase, is postulated to be an important source of cADPR in vivo. Although an increase in [Ca^<2+>]i is a key signal neutrophil functions, the mechanisms for regulation of [Ca^<2+>]i is unclear. The present study examined the regulation by cADPR of Ca^<2+> dynamics and its physiological role in human neutrophils.cADPR induced Ca^<2+> release from digitonin-permeabilized neutrophils and the release was blocked by 8Br-cADPR, an antagonist of cADPR and FK506 and rapamycin. fMLP and PAF induced a initial rapid rise of [Ca^<2+>]i and the following sustained rise in intact neutrophils. β-NAD^+ added into the medium increased [Ca^<2+>]i. Chemoattractans- and β-NAD^+-induced [Ca^<2+>]i rise were reduced by 8Br-cADPR, anti-CD38 antibody, FK506, rapamycin, NAD … More ase and several nucleoside transporter (NT) inhibitors. ENT_1, ENT_2, CNT_2, CNT_3 are expressed in neutrophils. These results suggest that cADPR synthesized extracellulary by CD38 transported into the cells through NTs and mobilize Ca^<2+> by FK506-binding protein-dependent process and is required for sustained Ca^<2+> influx in neutrophils.Pretreatment of human neutrophils with either EGTA or 8Br-cADPR specifically blocked the chemotsxis stimulated with fMLP or PAF. Likewise, treatment of neutrophils with FK506, anti-CD38 antibody, NADase and NT inhibitors blocked the chemotaxis to fMLP or PAF. These results demonstrate that neutrophil chemotaxis to fMLP and PAF are dependent on Ca^<2+> mobilization mediated by cADPR.Thus, cADPR controls neutrophil chemotaxis to chemoattractants through its production of cADPR, and acts as a critical regulator of inflammation and innate immune responses. Since many of the chemoattractant receptors regulated by cADPR bind to ligands that are associated with clinical pathology, cADPR, CD38 and NTs represent novel drug targets with potential application in chronic inflammatory and neurodegenerative disease. Less

环状ADP-ribose（CADPR）具有强大的Ca^<2+> - 动员动作，并作为许多激动剂的第二使者，从而调节了大量的Ca^<2+> - 介导的细胞过程。 CD38是特征最佳的哺乳动物ADP-核糖基环化酶，被张贴为体内CADPR的重要来源。尽管[Ca^<2+>] i的增加是钥匙信号中性粒细胞函数，但调节[Ca^<2+>] i的机制尚不清楚。本研究检查了CADPR对CA^<2+>动力学的调节及其在人类嗜中性粒细胞中的身体作用。CADPR诱导的Ca^<2+>从digitonin-渗透性中性粒细胞中释放出来，并且该释放被8BR-CADPR阻塞，8BR-CADPR是CADPR和FK506和FK506和RAPAMYCIN的拮抗剂。 FMLP和PAF诱导[Ca^<2+>] I的初始快速上升，并且完整的中性粒细胞的持续增长。将β-NAD^+添加到培养基中增加了[Ca^<2+>] i。 8BR-CADPR，抗CD38抗体，FK506，Rapamycin，NAD…更多ASE和几个核外侧转运蛋白（NT）抑制剂，将chemoattracttractans-和β-NAD^+ - 诱导的[Ca^<2+>] I升高降低。 ENT_1，ENT_2，CNT_2，CNT_3在中性粒细胞中表达。这些结果表明，CADPR通过CD38通过NTS运输到细胞的细胞外合成，并通过FK506结合蛋白依赖性过程动员Ca^<2+>，并且需要持续的ca^<2+>涌入中性粒细胞中的中性粒细胞中嗜中性粒细胞中的中性粒细胞中的中性粒细胞与EGTA或8BR-CADPRETER构成FMSIS的pRETSIS。同样，用FK506，抗CD38抗体，NADase和NT抑制剂治疗中性粒细胞将趋化性阻断了FMLP或PAF。这些结果表明，对FMLP和PAF的中性粒细胞趋化性取决于CADPR.CADPR介导的CA^<2+>动员，CADPR控制中性粒细胞通过产生CADPR的趋化剂，并通过其关键的炎症调节剂和与世的免疫反应。由于CADPR调节的许多趋化受体与与临床病理学有关的配体结合，CADPR，CD38和NTS代表了具有潜在应用于慢性炎症性和神经退行性疾病的新型药物靶标。较少的