PEP-1-SIRT2 inhibits inflammatory response and oxidative stress-induced cell death via expression of antioxidant enzymes in murine macrophages

Free Radic Biol Med. 2013 Oct:63:432-45. doi: 10.1016/j.freeradbiomed.2013.06.005. Epub 2013 Jun 12.

Abstract

Sirtuin 2 (SIRT2), a member of the sirtuin family of proteins, plays an important role in cell survival. However, the biological function of SIRT2 protein is unclear with respect to inflammation and oxidative stress. In this study, we examined the protective effects of SIRT2 on inflammation and oxidative stress-induced cell damage using a cell permeative PEP-1-SIRT2 protein. Purified PEP-1-SIRT2 was transduced into RAW 264.7 cells in a time- and dose-dependent manner and protected against lipopolysaccharide- and hydrogen peroxide (H₂O₂)-induced cell death and cytotoxicity. Also, transduced PEP-1-SIRT2 significantly inhibited the expression of cytokines as well as the activation of NF-κB and mitogen-activated protein kinases (MAPKs). In addition, PEP-1-SIRT2 decreased cellular levels of reactive oxygen species (ROS) and of cleaved caspase-3, whereas it elevated the expression of antioxidant enzymes such as MnSOD, catalase, and glutathione peroxidase. Furthermore, topical application of PEP-1-SIRT2 to 12-O-tetradecanoylphorbol 13-acetate-treated mouse ears markedly inhibited expression levels of COX-2 and proinflammatory cytokines as well as the activation of NF-κB and MAPKs. These results demonstrate that PEP-1-SIRT2 inhibits inflammation and oxidative stress by reducing the levels of expression of cytokines and ROS, suggesting that PEP-1-SIRT2 may be a potential therapeutic agent for various disorders related to ROS, including skin inflammation.

Keywords: Cell viability; Cytokines; Free radicals; Inflammation; PEP-1–SIRT2; Protein therapy; ROS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antioxidants / metabolism
  • Cell Death / drug effects
  • Cell Death / genetics
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Hydrogen Peroxide / toxicity
  • Inflammation / drug therapy
  • Inflammation / metabolism*
  • Inflammation / pathology
  • Lipopolysaccharides / toxicity
  • Macrophages / drug effects
  • Macrophages / enzymology
  • Macrophages / metabolism
  • Mice
  • Oxidative Stress*
  • Peptides / administration & dosage
  • Peptides / chemical synthesis
  • Peptides / genetics
  • Peptides / metabolism*
  • Phorbol Esters / toxicity
  • Reactive Oxygen Species / metabolism
  • Sirtuin 2 / administration & dosage
  • Sirtuin 2 / genetics
  • Sirtuin 2 / metabolism*
  • Skin / drug effects
  • Skin / growth & development

Substances

  • Antioxidants
  • Lipopolysaccharides
  • Peptides
  • Phorbol Esters
  • Reactive Oxygen Species
  • Hydrogen Peroxide
  • Sirt2 protein, mouse
  • Sirtuin 2