C-reactive protein induces matrix metalloproteinase-1 and -10 in human endothelial cells: implications for clinical and subclinical atherosclerosis.
OBJECTIVE
We examined the effect of C-reactive protein (CRP) on matrix metalloproteinase (MMP) and inhibitor expression in endothelial cells and in patients with clinical and subclinical atherosclerosis.


BACKGROUND
In addition to predicting atherosclerotic vascular disease, CRP may directly promote a proinflammatory/proatherosclerotic phenotype.


METHODS
Human umbilical vein endothelial cells (HUVECs) and aortic endothelial cells (HAECs) were incubated in the presence or absence of CRP (50 mug/ml). Microarray analysis, real-time polymerase chain reaction, immunological and activity assays for MMPs were performed. Specific inhibitors of mitogen-activated protein kinase pathway were used. The MMP-1 and -10 plasma levels were measured in apparently healthy subjects (n = 70). Immunolocalization of CRP, MMP-1, and MMP-10 was performed in human mammary arteries and carotid endarterectomy specimens.


RESULTS
C-reactive protein augmented MMP-1 and -10 messenger ribonucleic acid expression in HUVEC (p < 0.05) and HAEC (p < 0.01). C-reactive protein stimulation also increased MMP-1 and -10 protein in conditioned culture medium (p < 0.001), as well as MMP activity (p = 0.001). Specific inhibition of p38 or MEK abolished the CRP induction of the MMP-1, whereas MMP-10 induction blockade required the simultaneous inhibition of p38 and Jun N-terminal kinase pathways. Subjects with CRP values >3 mg/l (n = 37) had increased plasma MMP-1 and -10 (p < 0.05), the association being significant after adjustment for confounding variables (p = 0.04 and p = 0.008, respectively). The MMP-10 levels were elevated in subjects with higher carotid intima-media thickness (p = 0.009). Increased CRP and MMP-10 colocalized in endothelial layer and macrophage-rich areas in advanced atherosclerotic plaques.


CONCLUSIONS
Increased local and systemic CRP-related MMP activation might provide a link between inflammation and plaque vulnerability.
Laboratory of Atherosclerosis, School of Medicine/University Clinic, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Ines
I
Montero
Josune
J
Orbe
Nerea
N
Varo
Oscar
O
Beloqui
José I
JI
Monreal
José A
JA
Rodríguez
Javier
J
Díez
Peter
P
Libby
José A
JA
Páramo
D000328:Adult; D001011:Aorta / Q000166:cytology; D050197:Atherosclerosis / Q000209:etiology / Q000378:metabolism*; D002097:C-Reactive Protein / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D002339:Carotid Arteries / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D004790:Enzyme Induction / Q000187:drug effects; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D020935:MAP Kinase Signaling System / Q000502:physiology; D008297:Male; D008323:Mammary Arteries / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis* / Q000097:blood / Q000378:metabolism; D053506:Matrix Metalloproteinase 10; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000097:blood / Q000378:metabolism; D046228:Microarray Analysis; D008875:Middle Aged; D011994:Recombinant Proteins / Q000494:pharmacology; D013194:Staining and Labeling; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000097:blood; D014471:Umbilical Veins / Q000166:cytology
D011994:Recombinant Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D002097:C-Reactive Protein; D008666:Metalloendopeptidases; D053506:Matrix Metalloproteinase 10; D020781:Matrix Metalloproteinase 1