Nox4 and Duox1/2 Mediate Redox Activation of Mesenchymal Cell Migration by PDGF

Pyotr A Tyurin-Kuzmin; Nadezhda D Zhdanovskaya; Anna A Sukhova; George D Sagaradze; Eugene A Albert; Ludmila V Ageeva; George V Sharonov; Alexander V Vorotnikov; Vsevolod A Tkachuk

doi:10.1371/journal.pone.0154157

Nox4 and Duox1/2 Mediate Redox Activation of Mesenchymal Cell Migration by PDGF

PLoS One. 2016 Apr 25;11(4):e0154157. doi: 10.1371/journal.pone.0154157. eCollection 2016.

Authors

Affiliations

¹ Department of Biochemistry and Molecular Medicine, Faculty of Fundamental Medicine, M.V. Lomonosov Moscow State University, Moscow, Russia.
² Laboratory of Optical Microscopy and Spectroscopy of Biomolecules, Department of Structural Biology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
³ Institute of Experimental Cardiology, Russian Cardiology Research Center, Moscow, Russia.

Abstract

Platelet derived growth factor (PDGF) orchestrates wound healing and tissue regeneration by regulating recruitment of the precursor mesenchymal stromal cells (MSC) and fibroblasts. PDGF stimulates generation of hydrogen peroxide that is required for cell migration, but the sources and intracellular targets of H2O2 remain obscure. Here we demonstrate sustained live responses of H2O2 to PDGF and identify PKB/Akt, but not Erk1/2, as the target for redox regulation in cultured 3T3 fibroblasts and MSC. Apocynin, cell-permeable catalase and LY294002 inhibited PDGF-induced migration and mitotic activity of these cells indicating involvement of PI3-kinase pathway and H2O2. Real-time PCR revealed Nox4 and Duox1/2 as the potential sources of H2O2. Silencing of Duox1/2 in fibroblasts or Nox4 in MSC reduced PDGF-stimulated intracellular H2O2, PKB/Akt phosphorylation and migration, but had no such effect on Erk1/2. In contrast to PDGF, EGF failed to increase cytoplasmic H2O2, phosphorylation of PKB/Akt and migration of fibroblasts and MSC, confirming the critical impact of redox signaling. We conclude that PDGF-induced migration of mesenchymal cells requires Nox4 and Duox1/2 enzymes, which mediate redox-sensitive activation of PI3-kinase pathway and PKB/Akt.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetophenones / pharmacology
Animals
Cell Movement / drug effects
Chromones / pharmacology
Dual Oxidases
Gene Expression Regulation
HEK293 Cells
Humans
Hydrogen Peroxide / metabolism*
Mesenchymal Stem Cells / cytology
Mesenchymal Stem Cells / drug effects*
Mesenchymal Stem Cells / metabolism
Mice
Mitogen-Activated Protein Kinase 1 / genetics
Mitogen-Activated Protein Kinase 1 / metabolism
Mitogen-Activated Protein Kinase 3 / genetics
Mitogen-Activated Protein Kinase 3 / metabolism
Morpholines / pharmacology
NADPH Oxidase 4
NADPH Oxidases / antagonists & inhibitors
NADPH Oxidases / genetics*
NADPH Oxidases / metabolism
NIH 3T3 Cells
Oxidation-Reduction
Phosphatidylinositol 3-Kinase / genetics
Phosphatidylinositol 3-Kinase / metabolism
Platelet-Derived Growth Factor / antagonists & inhibitors
Platelet-Derived Growth Factor / pharmacology*
Primary Cell Culture
Proto-Oncogene Proteins c-akt / genetics
Proto-Oncogene Proteins c-akt / metabolism
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Signal Transduction

Substances

Acetophenones
Chromones
Morpholines
Platelet-Derived Growth Factor
RNA, Small Interfering
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
acetovanillone
Hydrogen Peroxide
Dual Oxidases
NADPH Oxidase 4
NADPH Oxidases
NOX4 protein, human
DUOX1 protein, human
DUOX2 protein, human
Phosphatidylinositol 3-Kinase
Proto-Oncogene Proteins c-akt
MAPK1 protein, human
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3

Grants and funding

Russian Science Foundation (http://рнф.рф/en/) grants 14-24-00086 (fibroblast migration and proliferation) and 14-35-00026 (microscopy and redox imaging) to VAT, Russian Foundation for Basic Research (http://www.rfbr.ru/rffi/eng) grant 14-04-01746a to AVV (redox signaling and PKB/Akt phosphorylation). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.