c-Jun N-terminal kinase and Akt signalling pathways regulating tumour necrosis factor-α-induced interleukin-32 expression in human lung fibroblasts: implications in airway inflammation
c-Jun N-terminal kinase and Akt signalling pathways regulating tumour necrosis factor-α-induced interleukin-32 expression in human lung fibroblasts: implications in airway inflammation
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DOI：
10.1111/imm.12374
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发表时间：
2015-02-01
期刊：
影响因子：
6.4
通讯作者：
Cao, Ju
中科院分区：
文献类型：
--
作者：
Li, Dagen;Chen, Dapeng;Cao, Ju
慢性阻塞性肺病 (COPD) 和哮喘等气道炎症性疾病与白细胞介素 32 (IL-32) 表达升高有关，白介素 32 是一种最近描述的细胞因子，似乎在炎症中发挥着关键作用。然而，迄今为止，肺部IL-32产生的调节尚未完全建立。我们检测了原代人肺成纤维细胞中肿瘤坏死因子 (TNF-) 的 IL-32 表达。在存在或不存在 TNF 和/或其他细胞因子/Toll 样受体 (TLR) 配体或各种信号分子抑制剂的情况下培养人肺成纤维细胞，通过定量 RT-PCR 和 ELISA 分析 IL-32 的表达。接下来，通过蛋白质印迹研究 Akt 和 c-Jun N 末端激酶 (JNK) 信号通路的激活。四种剪接异构体的白细胞介素 32 mRNA（、、和）在 TNF 刺激后上调，这与 TNF 激活的人肺成纤维细胞显着释放 IL-32 蛋白有关。干扰素和TNF的组合诱导增强人肺成纤维细胞中的IL-32释放，而IL-4、IL-17A、IL-27和TLR配体单独或与TNF-组合不改变人肺成纤维细胞中的IL-32释放。此外，Akt 和 JNK 通路的激活调节 TNF 诱导的人肺成纤维细胞中 IL-32 的表达，抑制 Akt 和 JNK 通路能够将 IL-32 的释放增加抑制到接近基础水平。这些数据表明 TNF-α 可能通过激活 Akt 和 JNK 信号通路诱导 IL-32 参与气道炎症。因此，TNF-/IL-32轴可能成为气道炎症性疾病的潜在治疗靶点。
Airway inflammatory diseases such as chronic obstructive pulmonary disease (COPD) and asthma are associated with elevated expression of interleukin-32 (IL-32), a recently described cytokine that appears to play a critical role in inflammation. However, so far, the regulation of pulmonary IL-32 production has not been fully established. We examined the expression of IL-32 by tumour necrosis factor- (TNF-) in primary human lung fibroblasts. Human lung fibroblasts were cultured in the presence or absence of TNF- and/or other cytokines/Toll-like receptor (TLR) ligands or various signalling molecule inhibitors to analyse the expression of IL-32 by quantitative RT-PCR and ELISA. Next, activation of Akt and c-Jun N-terminal kinase (JNK) signalling pathways was investigated by Western blot. Interleukin-32 mRNA of four spliced isoforms (, , and ) was up-regulated upon TNF- stimulation, which was associated with a significant IL-32 protein release from TNF--activated human lung fibroblasts. The combination of interferon- and TNF- induced enhanced IL-32 release in human lung fibroblasts, whereas IL-4, IL-17A, IL-27 and TLR ligands did not alter IL-32 release in human lung fibroblasts either alone, or in combination with TNF-. Furthermore, the activation of Akt and JNK pathways regulated TNF--induced IL-32 expression in human lung fibroblasts, and inhibition of the Akt and JNK pathways was able to suppress the increased release of IL-32 to nearly the basal level. These data suggest that TNF- may be involved in airway inflammation via the induction of IL-32 by activating Akt and JNK signalling pathways. Therefore, the TNF-/IL-32 axis may be a potential therapeutic target for airway inflammatory diseases.