Spexin Diminishes Atrial Fibrillation Vulnerability by Acting on Galanin Receptor 2.

Abstract

G protein-coupled receptors play a critical role in atrial fibrillation (AF). Spexin is a novel ligand of galanin receptors (GALRs). In this study, we investigated the regulation of spexin and GALRs on AF and the underlying mechanisms. Global spexin knockout (SPX-KO) and cardiomyocyte-specific GALRs knockout (GALR-cKO) mice underwent burst pacing electrical stimulation. Optical mapping was used to determine atrial conduction velocity and action potential duration. Atrial myocyte action potential duration and inward rectifying K⁺ current (<i>I</i>_K1) were recorded using whole-cell patch clamps. Isolated cardiomyocytes were stained with Fluo-3/AM dye, and intracellular Ca²⁺ handling was examined by CCD camera. A mouse model of AF was established by Ang-II (angiotensin II) infusion. Spexin plasma levels in patients with AF were lower than those in subjects without AF, and knockout of spexin increased AF susceptibility in mice. In the atrium of SPX-KO mice, potassium inwardly rectifying channel subfamily J member 2 (<i>KCNJ2</i>) and sarcolipin (<i>SLN</i>) were upregulated; meanwhile, <i>I</i>_K1 current was increased and Ca²⁺ handling was impaired in isolated atrial myocytes of SPX-KO mice. GALR2-cKO mice, but not GALR1-cKO and GALR3-cKO mice, had a higher incidence of AF, which was associated with higher <i>I</i>_K1 current and intracellular Ca²⁺ overload. The phosphorylation level of CREB (cyclic AMP responsive element binding protein 1) was upregulated in atrial tissues of SPX-KO and GALR2-cKO mice. Chromatin immunoprecipitation confirmed the recruitment of p-CREB to the proximal promoter regions of <i>KCNJ2</i> and <i>SLN</i>. Finally, spexin treatment suppressed CREB signaling, decreased <i>I</i>_K1 current and decreased intracellular Ca²⁺ overload, which thus reduced the inducibility of AF in Ang-II-infused mice. Spexin reduces atrial fibrillation susceptibility by inhibiting CREB phosphorylation and thus downregulating <i>KCNJ2</i> and <i>SLN</i> transcription by GALR2 receptor. The spexin/GALR2/CREB signaling pathway represents a novel therapeutic avenue in the development of agents against atrial fibrillation.