Meakins-Christie Laboratories, McGill University Heath Centre Research Institute, 3626 St-Urbain, Montréal, H2X 2P2, Canada.
J Cyst Fibros. 2011 Mar;10(2):107-13. doi: 10.1016/j.jcf.2010.11.005. Epub 2010 Dec 21.
Understanding the mechanisms underlying bacterial-driven inflammation and neutrophil recruitment is important to design better therapies for CF. CXCL8 is an important chemokine found elevated in the airways of CF patients that recruits neutrophil to sites of the inflammation.
Airway epithelial cells (AECs) expressing wild-type CFTR or CFTR∆F508 were challenged with Pseudomonas aeruginosa diffusible material (PsaDM) and the synthesis of CXCL8 was measured by quantitative real-time PCR and ELISA in absence or presence of MAPK inhibitors, TLR antagonists, glutathione and a NADPH oxidase inhibitor.
CFTR∆F508 AECs secrete more CXCL8 in response to PsaDM than their wild type counterpart, which can be reversed by addition of extracellular glutathione or incubating AECs at 27°C to favour folding and expression of CFTR at the cell membrane. Moreover, in CFTR∆F508 AECs, TLR2, TLR4 and TLR5 act redundantly to drive CXCL8 synthesis via the activation of NADPH oxidase.
These results demonstrate that NADPH oxidase is necessary for CXCL8 synthesis in response to TLRs activation by P. aeruginosa.
了解细菌驱动的炎症和中性粒细胞募集的机制对于设计更好的 CF 治疗方法非常重要。CXCL8 是一种在 CF 患者气道中升高的重要趋化因子，可将中性粒细胞募集到炎症部位。
用铜绿假单胞菌可扩散物质（PsaDM）刺激表达野生型 CFTR 或 CFTR∆F508 的气道上皮细胞（AECs），并通过定量实时 PCR 和 ELISA 测量 CXCL8 的合成，在不存在或存在 MAPK 抑制剂、TLR 拮抗剂、谷胱甘肽和 NADPH 氧化酶抑制剂的情况下。
CFTR∆F508 AECs 分泌更多的 CXCL8 以响应 PsaDM 比它们的野生型对应物，这可以通过添加细胞外谷胱甘肽或在 27°C 下孵育 AECs 来逆转，以有利于 CFTR 在细胞膜上的折叠和表达。此外，在 CFTR∆F508 AECs 中，TLR2、TLR4 和 TLR5 通过激活 NADPH 氧化酶冗余地作用以驱动 CXCL8 的合成。
这些结果表明，NADPH 氧化酶对于对 P. aeruginosa 激活的 TLRs 反应中的 CXCL8 合成是必要的。
