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Direct endothelial cell toxicity caused by C‐reactive protein: Role of oxidant mediated apoptosis
Author(s) -
Joshi Mandar S,
Tong Liyue,
Schanbacher Brandon L,
Bauer John Anthony
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1143
Subject(s) - apoptosis , toxicity , intracellular , viability assay , necrosis , programmed cell death , reactive oxygen species , pro oxidant , tumor necrosis factor alpha , chemistry , endothelial stem cell , pharmacology , biology , microbiology and biotechnology , biochemistry , medicine , endocrinology , in vitro
Elevation of plasma C‐reactive protein (CRP) and vascular endothelial injury/dysfunction have been separately recognized as important risk factors for coronary heart disease (CHD). Their interactions and contribution to disease progression are not clearly defined. We tested the hypothesis that clinically relevant concentrations of recombinant human CRP (rhCRP) were toxic to human endothelial cells (ECs), and that the toxicity was related to oxidant mediated cell apoptosis. ECs were incubated with rhCRP, or heat denatured rhCRP (0–10μg/ml, 48h). Cell survival, apoptosis & necrosis markers, and intracellular oxidant production were studied. Time and concentration dependent reductions in EC viability were observed for rhCRP (not hdCRP) and the primary mechanism was cellular apoptosis (% apoptosis 15.2±5.0; 36.01±2.7; 66.2±5.3: 0; 5; 10μg/ml CRP: p<0.05) rather than necrosis (<5% ‐ all conditions). Evidence of apoptosis was preceded by increased intracellular oxidant production (DCF intensity: 25283.53±4365.18; 24215.75±4796.25: 5; 10μg/ml CRP: p<0.05). Incubation with rhCRP in the presence of antioxidants (ascorbate or NAC) enhanced EC survival, reduced oxidant prevalence and prevented apoptosis. These data demonstrate that rhCRP causes direct EC toxicity and is mediated by oxidant related EC apoptosis. These observations may support the important predictive value of CRP as a CHD risk factor