Isoeugenol destabilizes IL-8 mRNA expression in THP-1 cells through induction of the negative regulator of mRNA stability tristetraprolin

Arch Toxicol. 2012 Feb;86(2):239-48. doi: 10.1007/s00204-011-0758-2. Epub 2011 Oct 4.

Abstract

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 μg/ml (0.61 mM) for isoeugenol, 100 μg/ml (0.58 mM) for DEM, 3 μg/ml (14.8 μM) for DNCB, and 250 μg/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 isoeugenol-treated 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.

MeSH terms

  • Allergens / pharmacology*
  • Cell Line
  • Cytokines / metabolism
  • Dinitrochlorobenzene / pharmacology
  • Eugenol / analogs & derivatives*
  • Eugenol / pharmacology
  • Gene Expression Regulation / drug effects*
  • Humans
  • Interleukin-8 / biosynthesis*
  • Interleukin-8 / genetics
  • Monocytes / drug effects*
  • Monocytes / metabolism
  • RNA Stability / drug effects*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Suppressor of Cytokine Signaling 3 Protein
  • Suppressor of Cytokine Signaling Proteins / metabolism
  • Tristetraprolin / biosynthesis*
  • Tristetraprolin / genetics
  • Tristetraprolin / metabolism

Substances

  • Allergens
  • Cytokines
  • Dinitrochlorobenzene
  • Interleukin-8
  • RNA, Messenger
  • SOCS3 protein, human
  • Suppressor of Cytokine Signaling 3 Protein
  • Suppressor of Cytokine Signaling Proteins
  • Tristetraprolin
  • Eugenol
  • isoeugenol