The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells

Qi bing Liu, Lu lu Liu, Ying mei Lu, Rong rong Tao, Ji yun Huang, Norifumi Shioda, Shigeki Moriguchi, Kohji Fukunaga, Feng Han, Yi jia Lou

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)

Abstract

The pathophysiological relevance of S-nitrosoglutathione (GSNO)-induced endothelial cell injury remains unclear. The main objective of this study was to elucidate the molecular mechanisms of GSNO-induced oxidative stress in endothelial cells. Morphological evaluation through DAPI staining and propidium iodide (PI) flow cytometry was used to detect apoptosis. In cultured EA.hy926 endothelial cells, exposure to GSNO led to a time- and dose-dependent apoptotic cascade. When intracellular reactive oxygen species (ROS) production was measured in GSNO-treated cells with the fluorescent probes 5-(and-6)-carboxy-2′,7′-dichlorofluorescein diacetate, we observed elevated ROS levels and a concomitant loss in mitochondrial membrane potential, indicating that GSNO-induced death signaling is mediated through a ROS-mitochondrial pathway. Importantly, we found that peroxynitrite formation and Omi/HtrA2 release from mitochondria were involved in this phenomenon, whereas changes of death-receptor dependent signaling were not detected in the same context. The inhibition of NADPH oxidase activation and Omi/HtrA2 by a pharmacological approach provided significant protection against caspase-3 activation and GSNO-induced cell death, confirming that GSNO triggers the death cascade in endothelial cells in a mitochondria-dependent manner. Taken together, our results indicate that ROS overproduction and loss of mitochondrial Omi/HtrA2 play a pivotal role in reactive nitrogen species-induced cell death, and the modulation of these pathways can be of significant therapeutic benefit.

Original languageEnglish
Pages (from-to)374-384
Number of pages11
JournalToxicology and Applied Pharmacology
Volume244
Issue number3
DOIs
Publication statusPublished - 2010 May

Keywords

  • Apoptosis
  • Eendothelial cells
  • Mitochondria
  • Peroxynitrite, Omi/HtrA2
  • S-nitrosoglutathione

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

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