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AMPK-Activated Protein Kinase Suppresses Ccr2 Expression by Inhibiting the NF-κB Pathway in RAW264.7 Macrophages

Fumiaki Kumase1, Kimio Takeuchi1, Yuki Morizane1,2

  • 1Retina Service, Angiogenesis Laboratory, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States of America.

Plos One
|January 23, 2016
PubMed

Insights

AMP-activated protein kinase (AMPK) negatively regulates C-C chemokine receptor 2 (Ccr2) expression in macrophages. AMPK activation reduces Ccr2 levels, partly via the NF-κB pathway, impacting inflammation.

Area of Science:

  • Cellular and Molecular Immunology
  • Macrophage Biology
  • Inflammation Research

Background:

  • C-C chemokine receptor 2 (Ccr2) is a critical marker for pro-inflammatory M1 macrophage activation.
  • The precise regulatory mechanisms governing Ccr2 expression are not fully understood.
  • AMP-activated protein kinase (AMPK) plays a significant role in both energy homeostasis and inflammation.

Purpose of the Study:

  • To investigate the role of AMPK in regulating Ccr2 protein levels in RAW264.7 macrophages.
  • To determine AMPK's influence on Ccr2 expression in both resting (M0) and LPS-induced M1 states.

Main Methods:

  • Utilized siRNA to knockdown the AMPKα1 subunit in RAW264.7 macrophages.
  • Employed pharmacologic AMPK activation using A769662.
  • Assessed Ccr2 protein levels in M0 and M1 macrophages under various conditions.
  • Investigated the involvement of NF-κB and TNF-α pathways.

Main Results:

  • Knockdown of AMPKα1 increased Ccr2 levels in both M0 and M1 macrophages.
  • Pharmacologic activation of AMPK attenuated LPS-induced increases in Ccr2 expression.
  • AMPK downregulation's effect on Ccr2 was partially reversed by NF-κB inhibition.
  • TNF-α inhibition showed minimal impact on Ccr2 levels.

Conclusions:

  • AMPK acts as a negative regulator of Ccr2 expression in RAW264.7 macrophages.
  • AMPK's inhibitory effect on Ccr2 is, in part, mediated through the NF-κB pathway.
  • These findings elucidate a novel regulatory mechanism for Ccr2 in macrophage activation.

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