Copyright © 2006, European Society of Cardiology
Antioxidative treatment inhibits the release of thrombogenic tissue factor from irradiation- and cytokine-induced endothelial cells
aDepartment of Cardiology and Pneumology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany
bInstitute of Pharmacy, Free University of Berlin, Berlin, Germany
cDepartment of Radiation Oncology and Radiotherapy, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany
dDivision of Hematology and Medical Oncology, Department of Medicine, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY, USA
* Corresponding author. Medical Clinic II, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, D-12203 Berlin, Germany. Tel.: +49 30 8445 2362; fax: +49 30 8441 9991. Email address: ursula.rauch{at}charite.de
Objectives: The aim of this study was to investigate the effect of the antioxidants pyrrolidine dithiocarbamate (PDTC) and N-acetylcysteine (NAC) on the ionizing radiation (IR)- and tumor necrosis factor-
(TNF-) induced tissue factor (TF) expression and its release from human umbilical vein endothelial cells (HUVECs).
Methods: HUVECs were irradiated with a single dose of either 5 Gy or 10 Gy and stimulated with TNF- (10 ng/mL) in the presence or absence of PDTC and NAC, respectively. Quantitative real-time PCR, ELISA, and TF activity measurements were performed, including TF activity in the supernatant. Apoptosis was detected by flow cytometric active caspase-3 measurement and formation of reactive oxygen species (ROS) by chemiluminescence.
Results: We demonstrated a thus far uninvestigated persistent induction of TF expression in HUVECs after treatment with IR and TNF-. Combined stimulation with IR and TNF- led to an immense shedding of microparticle-associated TF which was positively correlated with apoptosis and ROS formation. Antioxidative pre-treatment reduced not only apoptosis and ROS formation, but also the release of thrombogenic microparticles.
Conclusions: Antioxidative treatment inhibited apoptosis and shedding of microparticles, thereby reducing thrombogenicity. Thus, antioxidants may help to prevent late thrombosis after antiproliferative treatment when used in combination with anticoagulants.
KEYWORDS Tissue factor; Cytokines; Apoptosis; Antioxidants; Pyrrolidine dithiocarbamate; N-acetylcysteine