β2-adrenergic stimulation induces interleukin-6 by increasing Arid5a, a stabilizer of mRNA, through cAMP/PKA/CREB pathway in cardiac fibroblasts

Shota Tanaka; Atsuki Imaeda; Kotaro Matsumoto; Makiko Maeda; Masanori Obana; Yasushi Fujio

doi:10.1002/prp2.590

β2-adrenergic stimulation induces interleukin-6 by increasing Arid5a, a stabilizer of mRNA, through cAMP/PKA/CREB pathway in cardiac fibroblasts

Pharmacol Res Perspect. 2020 Apr;8(2):e00590. doi: 10.1002/prp2.590.

Authors

Shota Tanaka¹, Atsuki Imaeda¹, Kotaro Matsumoto¹, Makiko Maeda², Masanori Obana¹, Yasushi Fujio^{1

2

3}

Affiliations

¹ Laboratory of Clinical Science and Biomedicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita City, Osaka, Japan.
² Project of Clinical Pharmacology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University, Suita City, Osaka, Japan.
³ Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Suita City, Osaka, Japan.

Abstract

Background and purpose: In cardiovascular diseases, cardiac fibroblasts (CFs) participate in the myocardial inflammation by producing pro-inflammatory cytokines, worsening the prognosis. β2-adrenergic receptor (AR) and β3AR are expressed in CFs, and β-adrenergic stimulation promotes CFs to produce pro-inflammatory cytokines. However, the mechanism of the expression of pro-inflammatory cytokines in response to β-adrenergic stimulation remains to be fully elucidated.

Experimental approach: CFs were isolated from adult wild-type or AT-rich interactive domain-containing protein 5A (Arid5a) knockout mice. The expression of mRNA was measured by real-time RT-PCR. Interleukin (IL)-6 protein was measured by ELISA. The activity of nuclear factor-κB (NF-κB) and cyclic AMP (cAMP) response element binding protein (CREB) was assessed by ELISA-like assay or Western blotting.

Key results: The β-adrenergic stimulation remarkably induced IL-6 mRNA and protein through β2AR in CFs. The activation of adenylate cyclase and the enhancement of intracellular cAMP resulted in the upregulation of IL-6 mRNA expression. The induction of IL-6 transcript by β2AR signaling was independent of NF-κB. Concomitant with IL-6, the expression of Arid5a, an IL-6 mRNA stabilizing factor, was enhanced by β2-adrenergic stimulation and by cAMP increase. Importantly, β2AR signaling-mediated IL-6 induction was suppressed in Arid5a knockout CFs. Finally, β2AR stimulation phosphorylated CREB via PKA pathway, and the activation of CREB was essential for the induction of Arid5a and IL-6 mRNA.

Conclusion and implications: β2-adrenergic stimulation post-transcriptionally upregulates the expression of IL-6 by the induction of Arid5a through cAMP/PKA/CREB pathway in adult CFs. β2AR/Arid5a/IL-6 axis could be a therapeutic target against cardiac inflammation.

Keywords: fibroblasts; interleukin-6; β adrenergic receptor.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adrenergic beta-Agonists / pharmacology
Animals
Cyclic AMP / metabolism*
Cyclic AMP Response Element-Binding Protein / metabolism*
Cyclic AMP-Dependent Protein Kinases / metabolism*
DNA-Binding Proteins / genetics*
Female
Fibroblasts / metabolism*
Interleukin-6 / genetics*
Isoproterenol / pharmacology
Male
Mice, Inbred C57BL
Mice, Knockout
Myocardium / cytology
RNA, Messenger / metabolism
Receptors, Adrenergic, beta-2 / metabolism*
Signal Transduction
Transcription Factor RelA / metabolism
Transcription Factors / genetics*

Substances

Adrenergic beta-Agonists
Arid5a protein, mouse
Creb1 protein, mouse
Cyclic AMP Response Element-Binding Protein
DNA-Binding Proteins
Interleukin-6
RNA, Messenger
Receptors, Adrenergic, beta-2
Transcription Factor RelA
Transcription Factors
interleukin-6, mouse
Cyclic AMP
Cyclic AMP-Dependent Protein Kinases
Isoproterenol