Inhibition of peptidylarginine deiminase alleviates LPS-induced pulmonary dysfunction and improves survival in a mouse model of lethal endotoxemia

Eur J Pharmacol. 2018 Aug 15;833:432-440. doi: 10.1016/j.ejphar.2018.07.005. Epub 2018 Jul 5.


Immune cell death caused by neutrophil extracellular traps (NETs), referred to as NETosis, can contribute to the pathogenesis of endotoxemia and organ damage. Although the mechanisms by which infection induces NETosis and how that leads to organ dysfunction remain largely unknown, NET formation is often found following citrullination of histone H3 (CitH3) by peptidylarginine deiminase (PAD). We hypothesized that lipopolysaccharide (LPS)-induced activation of PAD and subsequent CitH3-mediated NET formation increases endothelial permeability and pulmonary dysfunction and, therefore, that inhibition of PAD can mitigate damage and improve survival in lethal endotoxemia. Here, we showed that treatment with YW3-56, a PAD2/PAD4 inhibitor, significantly diminished PAD activation, blocked LPS-induced pulmonary vascular leakage, alleviated acute lung injury, and improved survival in a mouse model of lethal LPS-induced endotoxemia. We found CitH3 in the bloodstream 30 min after intraperitoneal injection of LPS (35 mg/kg) into mice. Additionally, CitH3 production was induced in cultured neutrophils exposed to LPS, and NETs derived from these LPS-treated neutrophils increased the permeability of endothelial cells. However, YW3-56 reduced CitH3 production and NET formation by neutrophils following LPS exposure. Moreover, treatment with YW3-56 decreased the levels of circulating CitH3 and abolished neutrophil activation and NET formation in the lungs of mice with endotoxemia. These data suggest a novel mechanism by which PAD-NET-CitH3 can play a pivotal role in pulmonary vascular dysfunction and the pathogenesis of lethal endotoxemia.

Keywords: Citrullinated histone H3; Lipopolysaccharide; Microvascular permeability; Neutrophil extracellular traps; Peptidylarginine deiminase.

MeSH terms

  • 2-Naphthylamine / analogs & derivatives*
  • 2-Naphthylamine / pharmacology
  • 2-Naphthylamine / therapeutic use
  • Acute Lung Injury / drug therapy*
  • Acute Lung Injury / etiology
  • Acute Lung Injury / pathology
  • Animals
  • Arginine / analogs & derivatives*
  • Arginine / pharmacology
  • Arginine / therapeutic use
  • Citrullination / drug effects
  • Disease Models, Animal
  • Endothelial Cells / drug effects
  • Endothelial Cells / pathology
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology
  • Endotoxemia / chemically induced
  • Endotoxemia / complications
  • Endotoxemia / drug therapy*
  • Endotoxemia / mortality
  • Extracellular Traps / drug effects
  • Extracellular Traps / metabolism
  • Histones / metabolism*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Lipopolysaccharides / toxicity
  • Lung / blood supply
  • Lung / pathology
  • Mice
  • Mice, Inbred C57BL
  • Microvessels / cytology
  • Microvessels / drug effects
  • Microvessels / metabolism
  • Microvessels / pathology
  • Neutrophils / drug effects
  • Neutrophils / metabolism
  • Permeability / drug effects
  • Protein-Arginine Deiminases / antagonists & inhibitors*
  • Protein-Arginine Deiminases / metabolism
  • Treatment Outcome


  • Histones
  • Lipopolysaccharides
  • YW3-56
  • Arginine
  • 2-Naphthylamine
  • Protein-Arginine Deiminases