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Effects of H2O2 on Microfilaments of Cultured Endothelial Cells

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International Journal of Angiology

Abstract

Hydrogen peroxide (H2O2) is an important granulocyte derived mediator of endothelial cell injury. Alterations of the microfilaments system (especially of actin) in endothelial cells may be relevant for the pathogenesis of vascular leakage. In the study presented effects of H2O2 on actin monomers (G-actin) and filamentous actin (F-actin) were examined in cultured pulmonary artery endothelial cells. Phalloidin which blocks actin depolymerization by inhibiting actin monomer dissociation and C. Botulinum C2 toxin which ADP-ribosylates G-actin thereby inhibiting actin polymerization were used as tools for the study of H2O2–related actin alterations. Exposure of cells to 2 mM H2O2 resulted in a biphasic change of F-actin with an early decrease (15 min) and a subsequent doubling (120 min) paralleled by an inverse G-actin pattern. In endothelial cells with a 20% reduction of F-actin-brought about by preincubation with C2-toxin for 150 min- H2O2-related actin polymerization was unimpaired. In cells with completely dissolved F-actin (Bot. C2-toxin for 210 min) no actin polymerization occurred upon H2O2 application. Phalloidin blocked the early H2O2-induced F-actin decrease and slowed down late actin polymerization. Effects of H2O2 on endothelial actin were abolished in the presence of scavengers of oxygen metabolites (catalase) and by the poly (ADP) ribose-polymerase inhibitor aminobenzamid. The data presented are compatible with the concept that H2O2 stimulates actin turnover and actin nucleation which—in the long run—result in H2O2 related formation of new actin filaments. This process was blocked by oxygen metabolite scavengers and by inhibition of DNA strand break repair mechanisms.

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Acknowledgements

The authors thank Prof. Aktories, Department of Pharmacology and Toxicology, University Freiburg, for providing C. Botulinum C2-toxin. The technical assistance of S. Tannert-Otto is greatly appreciated. This study was supported by the Deutsche Forschungsgemeinschaft (SFB 547/B2 to N. Suttorp). N. Suttorp is a recipient of a Hermann and Lilly Schilling Professorship.

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Authors and Affiliations

  1. Department of Cardiovascular Surgery, Justus-Liebig University, Giessen, Germany

    Peter Roth M.D., F.I.C.A. & Friedrich-Wilhelm Hehrlein M.D.

  2. Department of Internal Medicine, Justus-Liebig University, Giessen, Germany

    Joachim Seybold M.D. & Norbert Suttorp M.D

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  1. Peter Roth M.D., F.I.C.A.
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  2. Joachim Seybold M.D.
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  3. Friedrich-Wilhelm Hehrlein M.D.
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  4. Norbert Suttorp M.D
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Roth, P., Seybold, J., Hehrlein, FW. et al. Effects of H2O2 on Microfilaments of Cultured Endothelial Cells . International Journal of Angiology 12, 62–67 (2003). https://doi.org/10.1007/s00547-003-0684-0

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  • DOI: https://doi.org/10.1007/s00547-003-0684-0

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