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

Background and Purpose In cardiovascular diseases, cardiac fibroblasts (CFs) participate in the myocardial inflammation by producing pro-inflammatory cytokines, worsening the prognosis. beta 2-adrenergic receptor (AR) and beta 3AR are expressed in CFs, and beta-adrenergic stimulation promotes CFs to produce pro-inflammatory cytokines. However, the mechanism of the expression of pro-inflammatory cytokines in response to beta-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-kappa B (NF-kappa B) and cyclic AMP (cAMP) response element binding protein (CREB) was assessed by ELISA-like assay or Western blotting. Key Results The beta-adrenergic stimulation remarkably induced IL-6 mRNA and protein through beta 2AR 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 beta 2AR signaling was independent of NF-kappa B. Concomitant with IL-6, the expression of Arid5a, an IL-6 mRNA stabilizing factor, was enhanced by beta 2-adrenergic stimulation and by cAMP increase. Importantly, beta 2AR signaling-mediated IL-6 induction was suppressed in Arid5a knockout CFs. Finally, beta 2AR 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 beta 2-adrenergic stimulation post-transcriptionally upregulates the expression of IL-6 by the induction of Arid5a through cAMP/PKA/CREB pathway in adult CFs. beta 2AR/Arid5a/IL-6 axis could be a therapeutic target against cardiac inflammation.