ω-Alkynyl arachidonic acid promotes anti-inflammatory macrophage M2 polarization against acute myocardial infarction via regulating the cross-talk between PKM2, HIF-1α and iNOS

Abstract

Macrophage polarization determines the timing for the switch from the inflammation phase to the inflammation resolution phase after acute myocardial infarction. The aim of the present study was to investigate whether ω-alkynyl arachidonic acid could mitigate the inflammatory lipid mediators in the regulation of macrophage phenotypes and functions with a special regard to myocardial infarction. We initially discovered that ω-alkynyl arachidonic acid selectively suppressed the up-regulation of inducible nitric oxide synthase (iNOS) over cyclooxygenase-2 (COX-2) in LPS-stimulated macrophages. ω-Alkynyl arachidonic acid also reduced the expression of macrophage M1 biomarkers (e.g., TNF-α, CXCL10, iNOS and IL-6) but increased the expression of macrophage M2 biomarkers (e.g., IL-10 and arginase-1) in LPS-stimulated macrophages. Moreover, ω-alkynyl arachidonic acid markedly enhanced the phagocytotic activity of macrophages against fluorescently-labeled beads or apoptotic H9c2 cardiac cells. We further investigated the in vivo cardioprotective activities of ω-alkynyl arachidonic acid in a mouse model of myocardial infarction. ω-Alkynyl arachidonic acid indeed reduced infarct size, cardiac damage and the leakage of myocardial enzymes CK-MB. Mechanistic studies revealed that ω-alkynyl arachidonic acid suppressed the overexpression and nuclear translocation of glycolytic enzyme PKM2 in LPS-stimulated macrophages. Furthermore, co-immunoprecipitation assay suggested that ω-alkynyl arachidonic acid disrupted the interaction between PKM2 and HIF-1α. Consequently, ω-alkynyl arachidonic acid diminished HIF-1α binding to the HRE sequence in iNOS promoter in response to LPS stimulation. Collectively, ω-alkynyl arachidonic acid may promote the anti-inflammatory M2 polarization of macrophages in acute myocardial infarction via regulating the cross-talk between PKM2, HIF-1α and iNOS.

Keywords: HIF-1a; M2 macrophage polarization; Myocardial ischemia-reperfusion injury; PKM2; iNOS; ω-Alkynyl arachidonic acid.