Acute activation of endothelial AMPK surprisingly inhibits endothelium-dependent hyperpolarization-like relaxations in rat mesenteric arteries

Br J Pharmacol. 2019 Aug;176(16):2905-2921. doi: 10.1111/bph.14716. Epub 2019 Jul 4.

Abstract

Background and purpose: Endothelium-dependent hyperpolarizations (EDHs) contribute to the regulation of peripheral resistance. They are initiated through opening of endothelial calcium-activated potassium channels (KCa ); the potassium ions released then diffuse to the underlying smooth muscle cells, causing hyperpolarization and thus relaxation. The present study aimed to examine whether or not AMPK modulates EDH-like relaxations in rat mesenteric arteries.

Experimental approach: Arterial rings were isolated for isometric tension recording. AMPK activity and protein level were measured by ELISA and western blotting respectively.

Key results: The AMPK activator, AICAR, reduced ACh-induced EDH-like relaxations and increased AMPK activity in preparations with endothelium; these responses were prevented by compound C, an AMPK inhibitor. AICAR inhibited relaxations induced by SKA-31 (opener of endothelial KCa ) but did not affect potassium-induced, hyperpolarization-attributable relaxations or increase AMPK activity in preparations without endothelium. A769662, another AMPK activator, not only caused a similar inhibition of relaxations to ACh and SKA-31 in preparations with endothelium but also inhibited hyperpolarization-attributable relaxations and augmented AMPK activity in rings without endothelium. Protein levels of total AMPKα, AMPKα1, or AMPKβ1/2 were comparable between preparations with and without endothelium.

Conclusions and implications: Activation of endothelial AMPK, by either AICAR or A769662, acutely inhibits EDH-like relaxations of rat mesenteric arteries. Furthermore, A769662 inhibits signalling downstream of smooth muscle hyperpolarization. In view of the major blunting effect of AMPK activation on EDH-like relaxations, caution should be applied when administering therapeutic agents that activate AMPK in patients with endothelial dysfunction characterized by reduced production and/or bioavailability of NO.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / physiology*
  • Acetylcholine / pharmacology
  • Aminoimidazole Carboxamide / analogs & derivatives
  • Aminoimidazole Carboxamide / pharmacology
  • Animals
  • Biphenyl Compounds
  • Bradykinin / pharmacology
  • Coronary Vessels / drug effects
  • Coronary Vessels / physiology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / physiology*
  • In Vitro Techniques
  • Male
  • Mesenteric Arteries / drug effects
  • Mesenteric Arteries / physiology*
  • Mice, Inbred C57BL
  • Pyrones / pharmacology
  • Rats, Sprague-Dawley
  • Ribonucleotides / pharmacology
  • Swine
  • Thiophenes / pharmacology
  • Vasodilation / drug effects

Substances

  • Biphenyl Compounds
  • Pyrones
  • Ribonucleotides
  • Thiophenes
  • Aminoimidazole Carboxamide
  • AMP-Activated Protein Kinases
  • AICA ribonucleotide
  • Acetylcholine
  • 4-hydroxy-3-(4-(2-hydroxyphenyl)phenyl)-6-oxo-7H-thieno(2,3-b)pyridine-5-carbonitrile
  • Bradykinin