Pirfenidone inhibits macrophage infiltration in 5/6 nephrectomized rats

Am J Physiol Renal Physiol. 2013 Mar 15;304(6):F676-85. doi: 10.1152/ajprenal.00507.2012. Epub 2012 Nov 14.

Abstract

Tubulointerstitial macrophage infiltration is a hallmark of chronic kidney disease involved in the progression of renal fibrosis. Pirfenidone is a newly identified antifibrotic drug, the potential mechanism of which remains unclear. The aim of this study was to investigate the effects of pirfenidone on M1/M2 macrophage infiltration in nephrectomized rats. Nephrectomized rats were treated with pirfenidone by gavage for 12 wk. Twenty-four hour urinary protein, N-acetyl-β-D-glycosaminidase (NAG) activity, systolic blood pressure, and C-reactive protein were determined. Paraffin-embedded sections were stained for CD68, CCR7, and CD163 macrophages. Monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1α (MIP-1α), as well as M1 and M2 macrophages secretory markers, were evaluated by real-time RT-PCR and Western blotting analysis. Pirfenidone significantly improved the elevated proteinuria and NAG activity from week 2 onward after surgery. Pirfenidone attenuated interstitial fibrosis and decreased expression of fibrotic markers including transforming growth factor-β(1), connective tissue growth factor, α-smooth muscle actin, fibronectin, and fibroblast-specific protein-1. Pirfenidone significantly decreased the infiltrating macrophages. The number of M1 and M2 macrophages was significantly lower after pirfenidone treatment. MCP-1 and MIP-1α were increased in nephrectomized rats at mRNA and protein levels. Pirfenidone treatment significantly inhibited their expression. The TNF-α, IL-6, and nitric oxide synthases-2 expressed by M1 macrophages were increased in nephrectomized rats, and pirfenidone significantly attenuated their expression. Pirfenidone treatment also significantly decreased arginase-1, dectin-1, CD206, and CD86 expressed by M2 macrophages. Thus pirfenidone inhibits M1 and M2 macrophage infiltration in 5/6 nephrectomized rats, which suggests its efficacy in the early and late periods of renal fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosaminidase / metabolism
  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology
  • Anti-Inflammatory Agents, Non-Steroidal / therapeutic use*
  • Biomarkers / metabolism
  • Blood Pressure / drug effects
  • Chemokine CCL2 / metabolism
  • Chemokine CCL3 / metabolism
  • Drug Evaluation, Preclinical
  • Kidney / drug effects
  • Kidney / metabolism
  • Kidney / pathology
  • Macrophages / drug effects*
  • Male
  • Nephrectomy
  • Nephrosclerosis / drug therapy*
  • Nephrosclerosis / immunology
  • Phenotype
  • Proteinuria / drug therapy
  • Pyridones / pharmacology
  • Pyridones / therapeutic use*
  • Rats
  • Rats, Sprague-Dawley
  • Renal Insufficiency, Chronic / drug therapy*
  • Renal Insufficiency, Chronic / enzymology
  • Renal Insufficiency, Chronic / immunology
  • Renal Insufficiency, Chronic / pathology

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Biomarkers
  • Ccl2 protein, rat
  • Chemokine CCL2
  • Chemokine CCL3
  • Pyridones
  • pirfenidone
  • Acetylglucosaminidase