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Forskolin attenuates the NLRP3 inflammasome activation and IL-1β secretion in human macrophages

Abstract

Background

The treatment of nucleotide-binding domain and leucine-rich repeat containing family, pyrin domain containing 3 (NLRP3) inflammasome-mediated pediatric inflammatory diseases is challenging. Here we studied whether cyclic adenosine monophosphate (cAMP) elevator forskolin could attenuate the nigericin-induced NLRP3-inflammasome activation and interleukin-1β (IL-1β) secretion in human macrophages.

Methods

The proteins and messenger RNA (mRNA) levels of inflammasome structural proteins and proinflammatory cytokines were measured in forskolin-stimulated nigericin-activated human THP-1 macrophages and primary macrophages.

Results

Activation of THP-1 macrophages with nigericin increased the mRNA expression of NLRP3, IL-1β, and caspase-1 (P < 0.01). Forskolin stimulation had no effect on the mRNA expression of NLRP3, caspase-1, or IL-1β in nigericin-activated cells (P > 0.05), while their protein levels were significantly decreased (P < 0.05). Forskolin-mediated increase in cytoplasmic cAMP in non-activated cells was attenuated in nigericin-activated macrophages (P < 0.05). Basal IL-1β secretion increased from 584 to 2696 pg/mL (P < 0.01) in nigericin-activated macrophages; forskolin dose-dependently reduced the nigericin-induced secretion of mature IL-1β (P < 0.01). Forskolin also inhibited the IL-1β secretion from activated human primary macrophages.

Conclusions

Forskolin inhibits the NLRP3 inflammasome activation and the secretion of mature IL-1β, in human macrophages. Forskolin and other cAMP elevator drugs could represent a novel approach for treatment of diseases associated with excessive inflammasome activation, like pediatric inflammatory diseases.

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Acknowledgements

We thank Xiaoping Shang at the Medical Records Department for the statistical assistance. This work was supported by the National Natural Science Foundation of China (81500588) and an independent research grant from the Finnish Cultural Foundation (00130167). The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.

Author information

Y.C. got the financial supports from the Foundations, conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures, reviewed drafts of the paper, redid the experiments, and revised the paper. J.G.W. performed the experiments, analyzed the data, wrote the paper, reviewed drafts of the paper, and revised the paper. J.J.F. performed the experiments, analyzed the data, prepared figures, reviewed drafts of the paper, and revised the paper. Y.H.W. performed the experiments, reviewed drafts of the paper, and revised the paper. T.F.L. contributed reagents and some materials, reviewed the draft of the paper, and revised the paper. K.N. contributed analysis protocols, helped to design the experiments, reviewed the manuscript, and revised the paper. K.K.E. contributed reagents, helped to design the experiments, reviewed the manuscript, and revised the paper. D.X. helped redo some experiments and reviewed the revised paper.

Competing interests

The authors declare no competing interests.

Correspondence to Yan Chen.

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