Netrin-1-treated macrophages protect the kidney against ischemia-reperfusion injury and suppress inflammation by inducing M2 polarization

Am J Physiol Renal Physiol. 2013 Apr 1;304(7):F948-57. doi: 10.1152/ajprenal.00580.2012. Epub 2013 Feb 13.


Improper macrophage activation is pathogenically linked to various metabolic, inflammatory, and immune disorders. Therefore, regulatory proteins controlling macrophage activation have emerged as important new therapeutic targets. We recently demonstrated that netrin-1 regulates inflammation and infiltration of monocytes and ameliorates ischemia-reperfusion-induced kidney injury. However, it was not known whether netrin-1 regulates the phenotype of macrophages and the signaling mechanism through which it might do this. In this study, we report novel mechanisms underlying netrin-1's effects on macrophages using in vivo and in vitro studies. Overexpression of netrin-1 in spleen and kidney of transgenic mice increased expression of arginase-1, IL-4, and IL-13 and decreased expression of COX-2, indicating a phenotypic switch in macrophage polarization toward an M2-like phenotype. Moreover, flow cytometry analysis showed a significant increase in mannose receptor-positive macrophages in spleen compared with wild type. In vitro, netrin-1 induced the expression of M2 marker expression in bone marrow-derived macrophages, peritoneal macrophages, and RAW264.7 cells, and suppressed IFNγ-induced M1 polarization and production of inflammatory mediators. Adoptive transfer of netrin-1-treated macrophages suppressed inflammation and kidney injury against ischemia-reperfusion. Netrin-1 activated PPAR pathways and inhibition of PPAR activation abolished netrin-1-induced M2 polarization and suppression of cytokine production. Consistent with in vitro studies, administration of PPAR antagonist to mice abolished the netrin-1 protective effects against ischemia-reperfusion injury of the kidney. These findings illustrate that netrin-1 regulates macrophage polarization through PPAR pathways and confers anti-inflammatory actions in inflammed kidney tissue.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anilides / pharmacology
  • Aniline Compounds / pharmacology
  • Animals
  • Inflammation / prevention & control
  • Interferon-gamma / pharmacology
  • Kidney / metabolism
  • Kidney / pathology*
  • Kidney Diseases / prevention & control
  • Macrophage Activation / drug effects
  • Macrophages, Peritoneal / drug effects*
  • Macrophages, Peritoneal / immunology
  • Male
  • Maleimides / pharmacology
  • Mice
  • Mice, Transgenic
  • Nerve Growth Factors / biosynthesis
  • Nerve Growth Factors / pharmacology*
  • Netrin-1
  • PPAR gamma / antagonists & inhibitors
  • PPAR-beta / antagonists & inhibitors
  • Reperfusion Injury / immunology
  • Reperfusion Injury / prevention & control*
  • Spleen / metabolism
  • Tumor Suppressor Proteins / biosynthesis
  • Tumor Suppressor Proteins / pharmacology*


  • 2-chloro-5-nitrobenzanilide
  • Anilides
  • Aniline Compounds
  • GSK 3987
  • Maleimides
  • Nerve Growth Factors
  • Ntn1 protein, mouse
  • PPAR gamma
  • PPAR-beta
  • Tumor Suppressor Proteins
  • Netrin-1
  • Interferon-gamma