Division of Cardiovascular Medicine, Department of Medical and Health Sciences, Faculty of Health Sciences, Linköping University, SE-581 85 Linköping, Sweden.
Eur J Cell Biol. 2010 Jun;89(6):462-7. doi: 10.1016/j.ejcb.2009.12.004. Epub 2010 Mar 5.
Adenosine deaminase (ADA) has a role in many immunity mediated disorders, such as asthma, tuberculosis and coronary artery disease. This study aims to investigate the ability of plasma ADA to modulate reactive oxygen species (ROS) production in neutrophils, and examine the involvement of adenosine and the cyclic AMP signaling pathway in this process.
Neutrophils were stimulated, in the absence or presence of plasma, with the chemotactic peptide fMLP (formyl-methionyl-leucyl-phenylalanine), and the ROS production was determined with luminol-enhanced chemiluminescence. Activity of ADA was measured spectrophotometrically.
Plasma dose-dependently amplified the ROS generation in fMLP-stimulated neutrophils. In parallel, incubation of neutrophils in plasma elevated the total ADA-activity approximately 10 times from 1.3 U/ml to 12 U/ml. Inhibition of ADA, or type IV phosphodiesterases, significantly lowered the plasma-mediated ROS production. Furthermore, the high-affinity adenosine A(1) receptor antagonists DPCPX and 8-phenyltheophylline markedly inhibited the plasma-induced respiratory burst in neutrophils, suggesting an A(1) receptor-mediated mechanism.
This study suggests that plasma ADA amplifies the release of toxic oxygen radicals from neutrophils through a downregulation of the inhibitory adenosine/cAMP-system and an enhanced activation of the stimulatory adenosine A(1)-receptor. This mechanism has probably a crucial role in regulating neutrophil function and in the defence against microbial infections. However, a sustained neutrophil activation could also contribute to inflammatory disorders such as atherosclerosis.
腺苷脱氨酶(ADA)在许多免疫介导的疾病中发挥作用,如哮喘、结核病和冠状动脉疾病。本研究旨在探讨血浆 ADA 调节中性粒细胞产生活性氧(ROS)的能力,并研究腺苷和环 AMP 信号通路在此过程中的作用。
在不存在或存在血浆的情况下,用趋化肽 fMLP(甲酰基-甲硫氨酸-亮氨酸-苯丙氨酸)刺激中性粒细胞,并通过发光增强化学发光法测定 ROS 的产生。用分光光度法测定 ADA 活性。
血浆剂量依赖性地放大了 fMLP 刺激的中性粒细胞中的 ROS 生成。同时,将中性粒细胞孵育在血浆中可使总 ADA 活性从 1.3 U/ml 升高约 10 倍至 12 U/ml。ADA 或 IV 型磷酸二酯酶的抑制显著降低了血浆介导的 ROS 产生。此外,高亲和力的腺苷 A1 受体拮抗剂 DPCPX 和 8-苯基茶碱显著抑制了血浆诱导的中性粒细胞呼吸爆发,表明存在 A1 受体介导的机制。
本研究表明,血浆 ADA 通过下调抑制性腺苷/cAMP 系统和增强刺激性腺苷 A1 受体的激活来放大中性粒细胞释放有毒氧自由基。这种机制可能在调节中性粒细胞功能和抵抗微生物感染方面起着至关重要的作用。然而,持续的中性粒细胞激活也可能导致炎症性疾病,如动脉粥样硬化。