First evidence for a crosstalk between mitochondrial and NADPH oxidase-derived reactive oxygen species in nitroglycerin-triggered vascular dysfunction

Antioxid Redox Signal. 2008 Aug;10(8):1435-47. doi: 10.1089/ars.2007.1969.


Chronic nitroglycerin treatment results in development of nitrate tolerance associated with endothelial dysfunction (ED). We sought to clarify how mitochondria- and NADPH oxidase (Nox)-derived reactive oxygen species (ROS) contribute to nitrate tolerance and nitroglycerin-induced ED. Nitrate tolerance was induced by nitroglycerin infusion in male Wistar rats (100 microg/h/4 day) and in C57/Bl6, p47(phox/) and gp91(phox/) mice (50 microg/h/4 day). Protein and mRNA expression of Nox subunits were unaltered by chronic nitroglycerin treatment. Oxidative stress was determined in vascular rings and mitochondrial fractions of nitroglycerin-treated animals by L-012 enhanced chemiluminescence, revealing a dominant role of mitochondria for nitrate tolerance development. Isometric tension studies revealed that genetic deletion or inhibition (apocynin, 0.35 mg/h/4 day) of Nox improved ED, whereas nitrate tolerance was unaltered. Vice versa, nitrate tolerance was attenuated by co-treatment with the respiratory chain complex I inhibitor rotenone (100 microg/h/4 day) or the mitochondrial permeability transition pore blocker cyclosporine A (50 microg/h/4 day). Both compounds improved ED, suggesting a link between mitochondrial and Nox-derived ROS. Mitochondrial respiratory chain-derived ROS are critical for the development of nitrate tolerance, whereas Nox-derived ROS mediate nitrate tolerance-associated ED. This suggests a crosstalk between mitochondrial and Nox-derived ROS with distinct mechanistic effects and sites for pharmacological intervention.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aorta / drug effects*
  • Aorta / metabolism
  • Aorta / physiopathology
  • Blotting, Western
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Cyclosporine / administration & dosage
  • Cyclosporine / pharmacology
  • Ethidium / analogs & derivatives
  • Ethidium / metabolism
  • Humans
  • In Vitro Techniques
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mitochondria, Heart / drug effects
  • Mitochondria, Heart / metabolism
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Nitroglycerin / administration & dosage
  • Nitroglycerin / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar
  • Reactive Nitrogen Species / metabolism
  • Reactive Oxygen Species / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Rotenone / administration & dosage
  • Rotenone / pharmacology
  • Transfection
  • Vasoconstriction / drug effects
  • Vasodilator Agents / administration & dosage
  • Vasodilator Agents / pharmacology


  • 2-hydroxyethidium
  • RNA, Messenger
  • Reactive Nitrogen Species
  • Reactive Oxygen Species
  • Vasodilator Agents
  • Rotenone
  • Cyclosporine
  • NADPH Oxidases
  • Ethidium
  • Nitroglycerin