Isoeugenol destabilizes IL-8 mRNA expression in THP-1 cells through induction of the negative regulator of mRNA stability tristetraprolin
Isoeugenol destabilizes IL-8 mRNA expression in THP-1 cells through induction of the negative regulator of mRNA stability tristetraprolin
复制标题
DOI：
10.1007/s00204-011-0758-2
复制
发表时间：
2012-02-01
影响因子：
6.1
通讯作者：
Corsini, Emanuela
中科院分区：
文献类型：
--
作者：
Galbiati, Valentina;Carne, Alice;Corsini, Emanuela
我们先前在人早幼粒细胞系THP-1中证明，除前半抗原异丁香酚外，所有测试的过敏原均诱导剂量相关的白细胞介素-8（IL-8）释放。在本研究中，我们研究了这种异常行为是否由富含AU的元素结合蛋白HuR和tristetraprolin（TTP）或下游细胞因子信号传导抑制分子（SOCS）-3调节。将接触性过敏原异丁香酚、马来酸二乙酯（DEM）和2，4-二硝基氯苯（DNCB）以及刺激性水杨酸用作参比化合物。用碘化丙啶染色法测定，化学品的浓度诱导细胞活力降低20%，即异丁香酚为100 μ g/ml（0.61mM），DEM为100 μ g/ml（0.58mM），DNCB为3 μ g/ml（14.8 μ M），水杨酸为250 μ g/ml（1.81mM）。IL-8 mRNA表达的时程实验和IL-8 mRNA半衰期的评估表明，异丁香酚处理的细胞中IL-8 mRNA稳定性降低。我们可以证明，在接触过敏原后，HuR和TTP的组合和调节导致IL-8 mRNA半衰期和释放的不同调节。用异丁香酚处理的THP-1细胞中TTP表达的增加导致IL-8 mRNA的不稳定，这可以解释IL-8释放的缺乏。相反，强过敏原DNCB不能上调TTP，而诱导HuR，导致IL-8 mRNA半衰期和蛋白质释放较长。SOCS-3仅在异丁香酚处理的细胞中被诱导;然而，其调节并没有拯救IL-8释放的缺乏，表明其不太可能参与IL-8产生的缺乏。最后，异丁香酚对IL-8 mRNA表达以及SOCS-3表达的去稳定作用导致抗炎作用，如异丁香酚调节LPS或离子霉素诱导的细胞因子释放的能力所证明的。
We previously demonstrated in the human promyelocytic cell line THP-1 that all allergens tested, with the exception of the prohapten isoeugenol, induced a dose-related release of interleukin-8 (IL-8). In the present study, we investigated whether this abnormal behavior was regulated by the AU-rich element-binding proteins HuR and tristetraprolin (TTP) or by the downstream molecule suppressor of cytokine signaling (SOCS)-3. The contact allergens isoeugenol, diethylmaleate (DEM), and 2,4-dinitrochlorobenzene (DNCB), and the irritant salicylic acid were used as reference compounds. Chemicals were used at concentrations that induced a 20% decrease in cell viability as assessed by propidium iodide staining, namely 100 mu g/ml (0.61 mM) for isoeugenol, 100 mu g/ml (0.58 mM) for DEM, 3 mu g/ml (14.8 mu M) for DNCB, and 250 mu g/ml (1.81 mM) for salicylic acid. Time course experiments of IL-8 mRNA expression and assessment of IL-8 mRNA half-life, indicated a decreased IL-8 mRNA stability in isoeugenoltreated cells. We could demonstrate that a combination and regulation of HuR and TTP following exposure to contact allergens resulted in a different modulation of IL-8 mRNA half-life and release. The increased expression of TTP in THP-1 cells treated with isoeugenol results in destabilization of the IL-8 mRNA, which can account for the lack of IL-8 release. In contrast, the strong allergen DNCB failing to up-regulate TTP, while inducing HuR, resulted in longer IL-8 mRNA half-life and protein release. SOCS-3 was induced only in isoeugenol-treated cells; however, its modulation did not rescue the lack of IL-8 release, indicating that it is unlikely to be involved in the lack of IL-8 production. Finally, the destabilization effect of isoeugenol on IL-8 mRNA expression together with SOCS-3 expression resulted in an anti-inflammatory effect, as demonstrated by the ability of isoeugenol to modulate LPS or ionomycin-induced cytokine release.