Heat shock-induced attenuation of hydroxyl radical generation and mitochondrial aconitase activity in cardiac H9c2 cells

Govindasamy Ilangovan, C. D. Venkatakrishnan, Anna Bratasz, Sola Osinbowale, Arturo J. Cardounel, Jay L. Zweier, Periannan Kuppusamy

Research output: Contribution to journalArticle

Abstract

A mild heat shock (hyperthermia) protects cells from apoptotic and necrotic deaths by inducing overexpression of various heat shock proteins (Hsps). These proteins, in combination with the activation of the nitric oxide synthase (NOS) enzyme, play important roles in the protection of the myocardium against a variety of diseases. In the present work we report that the generation of potent reactive oxygen species (ROS), namely ·OH in cardiac H9c2 cells, is attenuated by heat shock treatment (2 h at 42°C). Western blot analyses showed that heat shock treatment induced overexpression of Hsp70, Hsp60, and Hsp25. The observed ·OH was found to be derived from the superoxide (O2-.) generated by the mitochondria. Whereas the manganese superoxide dismutase (MnSOD) activity was increased in the heat-shocked cells, the mitochondrial aconitase activity was reduced. The mechanism of O2-. conversion into ·OH in mitochondria is proposed as follows. The O2-. leaked from the electron transport chain, oxidatively damages the mitochondrial aconitase, releasing a free Fe2+. The aconitase-released Fe2+ combines with H2O2 to generate ·OH via a Fenton reaction and the oxidized Fe3+ recombines with the inactivated enzyme after being reduced to Fe2+ by other cellular reductants, turning it over to be active. However, in heat-shocked cells, because of higher MnSOD activity, the excess H2O2 causes irreversible damage to the mitochondrial aconitase enzyme, thus inhibiting its activity. In conclusion, we propose that attenuation of ·OH generation after heat shock treatment might play an important role in reducing the myocardial ischemic injury, observed in heat shock-treated animals.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume290
Issue number2
DOIs
StatePublished - Feb 2006
Externally publishedYes

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Aconitate Hydratase
Hydroxyl Radical
Shock
Hot Temperature
Mitochondria
Superoxide Dismutase
Enzymes
Reducing Agents
Electron Transport
Heat-Shock Proteins
Nitric Oxide Synthase
Superoxides
Reactive Oxygen Species
Myocardium
Animals
Fever
Western Blotting
Chemical activation
Wounds and Injuries

Keywords

  • Free radicals
  • Proteins
  • Reactive oxygen species
  • Spin trapping

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Heat shock-induced attenuation of hydroxyl radical generation and mitochondrial aconitase activity in cardiac H9c2 cells. / Ilangovan, Govindasamy; Venkatakrishnan, C. D.; Bratasz, Anna; Osinbowale, Sola; Cardounel, Arturo J.; Zweier, Jay L.; Kuppusamy, Periannan.

In: American Journal of Physiology - Cell Physiology, Vol. 290, No. 2, 02.2006.

Research output: Contribution to journalArticle

Ilangovan, Govindasamy ; Venkatakrishnan, C. D. ; Bratasz, Anna ; Osinbowale, Sola ; Cardounel, Arturo J. ; Zweier, Jay L. ; Kuppusamy, Periannan. / Heat shock-induced attenuation of hydroxyl radical generation and mitochondrial aconitase activity in cardiac H9c2 cells. In: American Journal of Physiology - Cell Physiology. 2006 ; Vol. 290, No. 2.
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