IL-17A augments TNF-α-induced IL-6 expression in airway smooth muscle by enhancing mRNA stability
IL-17A augments TNF-α-induced IL-6 expression in airway smooth muscle by enhancing mRNA stability
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DOI：
10.1016/j.jaci.2004.06.023
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发表时间：
2004-10-01
影响因子：
14.2
通讯作者：
Ammit, AJ
中科院分区：
文献类型：
--
作者：
Henness, S;Johnson, CK;Ammit, AJ
背景：IL-17 A参与炎症调节，在哮喘气道中含量增加。人气道平滑肌（ASM）细胞合成细胞粘附分子和细胞因子以响应炎症介质目的：在这项研究中，我们研究了IL-17 A是否调节ASM细胞的合成功能方法：原代培养的ASM单独用IL-17 A处理，并与促炎细胞因子TNF-α和IL-1 β联合处理。ELISA法测定细胞间粘附分子1表达、GM-CSF和IL-6分泌。通过瞬时转染的启动子结构进行转录调控检查，而mRNA的稳定性进行了评估，放线菌素D追逐和定量real-time PCR.Results：气道平滑肌不分泌IL-17 A刺激后的ASM与TNF-α和IL-1 β。此外，IL-17 A（0.1-10 ng/mL）对TNF-α诱导和IL-1 β诱导的细胞间粘附分子1表达或GM-CSF分泌无影响。然而，IL-17 A（10 ng/mL）显著增加TNF-α诱导的IL-6分泌12倍（TNF-α，2.3 +/- 0.4 ng/mL; IL-17 A和TNF-α，27.5 +/- 4.8 ng/mL; P <0.05），而对IL-1 β诱导的IL-6无影响。虽然IL-17 A对TNF-α诱导的IL-6基因表达的核因子KB介导的转录调节没有影响，但IL-17 A显著增强TNF-α诱导的IL-6 mRNA的稳定性。总的来说，这些结果表明IL-17 A放大了ASM细胞的合成功能，通过转录后途径而不是转录机制起作用，增加TNF-α诱导的ASM细胞分泌IL-6。
Background: IL-17A is implicated in the regulation of inflammation and is found in increased amounts in the asthmatic airway. Human airway smooth muscle (ASM) cells synthesize cell adhesion molecules and cytokines in response to inflammatory mediators.Objective: In this study, we examined whether IL-17A modulated the synthetic function of ASM cells.Methods: Primary ASM cultures were treated with IL-17A alone and in combination with the proinflammatory cytokines TNF-alpha and IL-1beta. Intercellular adhesion molecule 1 expression, GM-CSF, and IL-6 secretion were measured by ELISA. Examination of transcriptional regulation was performed via transient transfection of promoter constructs, whereas mRNA stability was assessed by actinomycin D chase and quantitative real-time PCR.Results: Airway smooth muscle did not secrete IL-17A after stimulation of ASM with TNF-alpha and IL-1beta. Furthermore, IL-17A (0.1-10 ng/mL) had no effect on TNF-alpha-induced and IL-1beta-induced intercellular adhesion molecule 1 expression or GM-CSF secretion. However, IL-17A (10 ng/mL) significantly augmented TNF-alpha-induced IL-6 secretion 12-fold (TNF-alpha, 2.3 +/- 0.4 ng/mL; IL-17A and TNF-alpha, 27.5 +/- 4.8 ng/mL; P < .05) while having no effect on IL-1beta-induced IL-6. Although IL-17A had no effect on nuclear factor KB-mediated transcriptional regulation of IL-6 gene expression induced by TNF-alpha, IL-17A significantly augmented TNF-alpha-induced IL-6 mRNA stability.Conclusion: Collectively, these results demonstrate that IL-17A amplifies the synthetic function of ASM cells, acting via a posttranscriptional pathway, rather than transcriptional mechanisms, to augment TNF-alpha-induced secretion of IL-6 from ASM cells.