Dept. of Pharmacology, Chiesi Farmaceutici, Parma, Italy.
Am J Physiol Lung Cell Mol Physiol. 2012 Nov 15;303(10):L929-38. doi: 10.1152/ajplung.00046.2012. Epub 2012 Sep 14.
Interleukin-8 (IL-8/CXCL8) is an important neutrophil chemoattractant known to be elevated in the airways of cigarette smokers and in patients with chronic obstructive pulmonary disease (COPD). We examined the acute effect of aqueous cigarette smoke extract (CSE) on IL-8 expression in primary human pulmonary cells, in particular in normal human bronchial smooth muscle cells (HBSMCs). IL-8 mRNA levels increased upon CSE exposure in a concentration- and time-dependent manner, and such an effect was accompanied by IL-8 secretion. CSE-evoked elevation of IL-8 mRNA was mimicked by its component acrolein. Both CSE and acrolein induced p38 mitogen-activated protein kinase (MAPK) phosphorylation, accompanied by the phosphorylation of MAPK-activated kinase 2 (MK2), a known downstream substrate of the p38 MAPK, both in HBSMCs and in human airway epithelial cells. Furthermore, pharmacological inhibition of p38 MAPK or MK2 strongly accelerated the decay of IL-8 mRNA levels upon stimulation with CSE or acrolein and subsequent blockade of mRNA neosynthesis with actinomycin D in pulmonary structural cells (HBSMCs and airways epithelial cells) as well as in human alveolar macrophages. Conversely, pharmacological inhibition of ERK1/2 signaling inhibited CSE-induced steady-state levels of IL-8 mRNA without affecting mRNA stability, thus suggesting inhibition at the transcriptional level. In sum, p38 MAPK/MK2 signaling is an important posttranscriptional mechanism underlying upregulation of IL-8 mRNA levels elicited by CSE and acrolein. Given the pivotal role of IL-8 in neutrophil chemotaxis and activation, our results shed light on the mechanisms through which cigarette smoke can initiate inflammation in the lung.
白细胞介素-8(IL-8/CXCL8)是一种重要的中性粒细胞趋化因子,已知在吸烟人群的气道和慢性阻塞性肺疾病(COPD)患者中升高。我们研究了水相香烟烟雾提取物(CSE)对原代人肺细胞中 IL-8 表达的急性影响,特别是在正常的人支气管平滑肌细胞(HBSMCs)中。CSE 暴露后,IL-8 mRNA 水平呈浓度和时间依赖性增加,并且这种效应伴随着 IL-8 的分泌。CSE 成分丙烯醛模拟了 CSE 诱导的 IL-8 mRNA 升高。CSE 和丙烯醛均可诱导 p38 丝裂原活化蛋白激酶(MAPK)磷酸化,同时伴有 MAPK 激活激酶 2(MK2)磷酸化,这是 p38 MAPK 的已知下游底物,在 HBSMCs 和人气道上皮细胞中均如此。此外,在肺结构细胞(HBSMCs 和气道上皮细胞)以及人肺泡巨噬细胞中,用 p38 MAPK 或 MK2 的药理学抑制剂强烈抑制 CSE 或丙烯醛刺激后 IL-8 mRNA 水平的衰减,并随后用放线菌素 D 阻断 mRNA 新合成,可强烈抑制 CSE 或丙烯醛刺激后 IL-8 mRNA 水平的衰减,并随后用放线菌素 D 阻断 mRNA 新合成。相反,ERK1/2 信号通路的药理学抑制抑制了 CSE 诱导的 IL-8 mRNA 的稳态水平,而不影响 mRNA 的稳定性,因此提示在转录水平上的抑制。总之,p38 MAPK/MK2 信号通路是 CSE 和丙烯醛诱导 IL-8 mRNA 水平上调的重要转录后机制。鉴于 IL-8 在中性粒细胞趋化和激活中的关键作用,我们的结果揭示了香烟烟雾引发肺部炎症的机制。