C-reactive protein-induced in vitro endothelial cell activation is an artefact caused by azide and lipopolysaccharide

被引:135
作者
Taylor, KE
Giddings, JC
van den Berg, CW
机构
[1] Cardiff Univ, Wales Coll Med, Wales Heart Res Inst, Dept Pharmacol Therapeut & Toxicol, Cardiff CF14 4XN, S Glam, Wales
[2] Cardiff Univ, Wales Coll Med, Dept Haematol, Cardiff CF14 4XN, S Glam, Wales
关键词
C-reactive protein; endothelial cells; azide; LPS;
D O I
10.1161/01.ATV.0000164623.41250.28
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Objective - C- reactive protein ( CRP) has been proposed to be an independent risk factor for cardiovascular disease. In vitro studies investigating the mechanism behind this have used purified commercial CRP ( cCRP) and endothelial cells. We investigated the role of contaminants in cCRP preparations. Methods and Results - Human umbilical vein endothelial cells and the human endothelial cell line EA. hy926 were incubated with Escherichia coli - derived cCRP, in- house - generated azide- free recombinant, and ascites- purified CRP, azide, or lipopolysaccharide ( LPS) equivalent to the concentration present in cCRP preparations. Cells were investigated for change in cell proliferation, morphology, apoptosis, and expression of endothelial NO synthase and intercellular adhesion molecule- 1. Cell supernatants were assessed for monocyte chemoattractant protein- 1 ( MCP- 1), interleukin- 8, von Willebrand factor secretion, and pH change. Only cCRP was able to induce all activation events analyzed; however, this ability was lost on extensive dialysis, suggesting that low molecular weight contaminants were responsible for these events. Indeed, the effects of cCRP were mirrored by azide or LPS. Conclusions - We investigated a wide range of effects on endothelial cells ascribed to CRP; however, azide and LPS, but never CRP itself, were responsible for the cell activation events. We conclude that CRP, per se, does not activate endothelial cells.
引用
收藏
页码:1225 / 1230
页数:6
相关论文
共 35 条
[1]   Possible protective role for c-reactive protein in atherogenesis - Complement activation by modified lipoproteins halts before detrimental terminal sequence [J].
Bhakdi, S ;
Torzewski, M ;
Paprotka, K ;
Schmitt, S ;
Barsoom, H ;
Suriyaphol, P ;
Han, SR ;
Lackner, KJ ;
Husmann, M .
CIRCULATION, 2004, 109 (15) :1870-1876
[2]   Effects of C-reactive protein on the release of von Willebrand factor, E-selectin, thrombomodulin and intercellular adhesion molecule-1 from human umbilical vein endothelial cells [J].
Blann, AD ;
Lip, GYH .
BLOOD COAGULATION & FIBRINOLYSIS, 2003, 14 (04) :335-340
[3]   C-reactive protein induces apoptosis in human coronary vascular smooth muscle cells [J].
Blaschke, F ;
Bruemmer, D ;
Yin, F ;
Takata, Y ;
Wang, W ;
Fishbein, MC ;
Okura, T ;
Higaki, J ;
Graf, K ;
Fleck, E ;
Hsueh, WA ;
Law, RE .
CIRCULATION, 2004, 110 (05) :579-587
[4]   Effect of C-reactive protein on chemokine expression in human aortic endothelial cells [J].
Devaraj, S ;
Kumaresan, PR ;
Jialal, I .
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, 2004, 36 (03) :405-410
[5]   C-reactive protein binds to apoptotic cells, protects the cells from assembly of the terminal complement components, and sustains an antiinflammatory innate immune response: Implications for systemic autoimmunity [J].
Gershov, D ;
Kim, S ;
Brot, N ;
Elkon, KB .
JOURNAL OF EXPERIMENTAL MEDICINE, 2000, 192 (09) :1353-1363
[6]   Human C-reactive protein increases cerebral infarct size after middle cerebral artery occlusion in adult rats [J].
Gill, R ;
Kemp, JA ;
Sabin, C ;
Pepys, MB .
JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM, 2004, 24 (11) :1214-1218
[7]   C-reactive protein and complement are important mediators of tissue damage in acute myocardial infarction [J].
Griselli, M ;
Herbert, J ;
Hutchinson, WL ;
Taylor, KM ;
Sohail, M ;
Krausz, T ;
Pepys, MB .
JOURNAL OF EXPERIMENTAL MEDICINE, 1999, 190 (12) :1733-1739
[8]   C-reactive protein and cardiovascular disease: new insights from an old molecule [J].
Hirschfield, GM ;
Pepys, MB .
QJM-AN INTERNATIONAL JOURNAL OF MEDICINE, 2003, 96 (11) :793-807
[9]   C-reactive protein: Risk marker or mediator in atherothrombosis? [J].
Jialal, I ;
Devaraj, S ;
Venugopal, SK .
HYPERTENSION, 2004, 44 (01) :6-11
[10]   Conformational rearrangement in C-reactive protein is required for proinflammatory actions on human endothelial cells [J].
Khreiss, T ;
József, L ;
Potempa, LA ;
Filep, JG .
CIRCULATION, 2004, 109 (16) :2016-2022