AMP-Activated Protein Kinase alpha 2 in Neutrophils Regulates Vascular Repair via Hypoxia-Inducible Factor-1 alpha and a Network of Proteins Affecting Metabolism and Apoptosis

Abstract

Rationale: The AMP-activated protein kinase (AMPK) is stimulated by hypoxia, and although the AMPK alpha 1 catalytic subunit has been implicated in angiogenesis, little is known about the role played by the AMPK alpha 2 subunit in vascular repair. Objective: To determine the role of the AMPK alpha 2 subunit in vascular repair. Methods and Results: Recovery of blood flow after femoral artery ligation was impaired (>80%) in AMPK alpha 2(-/-) versus wild-type mice, a phenotype reproduced in mice lacking AMPK alpha 2 in myeloid cells (AMPK alpha 2(Delta MC)). Three days after ligation, neutrophil infiltration into ischemic limbs of AMPK alpha 2(Delta MC) mice was lower than that in wild-type mice despite being higher after 24 hours. Neutrophil survival in ischemic tissue is required to attract monocytes that contribute to the angiogenic response. Indeed, apoptosis was increased in hypoxic neutrophils from AMPK alpha 2(Delta MC) mice, fewer monocytes were recruited, and gene array analysis revealed attenuated expression of proangiogenic proteins in ischemic AMPK alpha 2(Delta MC) hindlimbs. Many angiogenic growth factors are regulated by hypoxia-inducible factor, and hypoxia-inducible factor-1 alpha induction was attenuated in AMPK alpha 2-deficient cells and accompanied by its enhanced hydroxylation. Also, fewer proteins were regulated by hypoxia in neutrophils from AMPK alpha 2(Delta MC) mice. Mechanistically, isocitrate dehydrogenase expression and the production of alpha-ketoglutarate, which negatively regulate hypoxia-inducible factor-1 alpha stability, were attenuated in neutrophils from wild-type mice but remained elevated in cells from AMPK alpha 2(Delta MC) mice. Conclusions: AMPK alpha 2 regulates alpha-ketoglutarate generation, hypoxia-inducible factor-1 alpha stability, and neutrophil survival, which in turn determine further myeloid cell recruitment and repair potential. The activation of AMPK alpha 2 in neutrophils is a decisive event in the initiation of vascular repair after ischemia