Implication of AMP-activated protein kinase in transient receptor potential vanilloid type 1-mediated activation of endothelial nitric oxide synthase

Mol Med. 2012;18:805-15. doi: 10.2119/molmed.2011.00461.

Abstract

We investigated whether AMP-activated protein kinase (AMPK), a multi-functional regulator of energy homeostasis, is involved in transient receptor potential vanilloid type 1 (TRPV1)-mediated activation of endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs) and mice. In ECs, treatment with evodiamine, the activator of TRPV1, increased the phosphorylation of AMPK, acetyl-CoA carboxylase (ACC) and eNOS, as revealed by western blot analysis. Inhibition of AMPK activation by compound C or dominant-negative AMPK mutant abrogated the evodiamine-induced increase in phosphorylation of AMPK and eNOS and NO bioavailability, as well as tube formation in ECs. Immunoprecipitation and two-hybrid analysis demonstrated that AMPK mediated the evodiamine-induced increase in the formation of a TRPV1-eNOS complex. Additionally, TRPV1 activation by evodiamine increased the phosphorylation of AMPK and eNOS in aortas of wild-type mice but did not activate eNOS in aortas of TRPV1-deficient mice. In mice, inhibition of AMPK activation by compound C markedly decreased evodiamine-evoked angiogenesis in Matrigel plugs and in a hind-limb ischemia model. Moreover, evodiamine-induced phosphorylation of AMPK and eNOS in aortas of apolipoprotein E deficient (ApoE(-/-)) mice was abrogated in TRPV1-deficient ApoE(-/-) mice. In conclusion, TRPV1 activation may trigger AMPK-dependent signaling, which leads to enhanced activation of AMPK and eNOS and retarded development of atherosclerosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / genetics
  • AMP-Activated Protein Kinases / metabolism*
  • Animals
  • Aorta / cytology
  • Aorta / metabolism*
  • Blood Vessels / drug effects
  • Blotting, Western
  • Cattle
  • Cells, Cultured
  • Collagen
  • Drug Combinations
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Enzyme Activation / drug effects
  • Hindlimb / blood supply
  • Hindlimb / metabolism
  • Injections, Intraperitoneal
  • Laminin
  • Male
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type III / genetics
  • Nitric Oxide Synthase Type III / metabolism*
  • Phosphorylation / drug effects
  • Protein Binding
  • Proteoglycans
  • Quinazolines / administration & dosage
  • Quinazolines / pharmacology
  • TRPV Cation Channels / genetics
  • TRPV Cation Channels / metabolism*

Substances

  • Drug Combinations
  • Laminin
  • Proteoglycans
  • Quinazolines
  • TRPV Cation Channels
  • TRPV1 protein, mouse
  • matrigel
  • Nitric Oxide
  • Collagen
  • evodiamine
  • Nitric Oxide Synthase Type III
  • AMP-Activated Protein Kinases