Immaturity of microvessels in haemorrhagic plaques is associated with proteolytic degradation of angiogenic factors.
OBJECTIVE
We investigated the causes of microvessel immaturity and destabilization in human atherosclerotic lesions.


RESULTS
Human atherosclerotic carotid plaques (n = 24) were classified as non-haemorrhagic (NH) or haemorrhagic (Hem), according to their macroscopic aspect and haemoglobin content. Plaque microvessel density and maturity were quantified by immunohistochemistry. Expression of angiogenic factors was studied by immunohistochemistry, in situ hybridization, and ELISA. Plaque-conditioned media were tested for plasmin and elastase activities and for their ability to degrade angiogenic factors and to induce smooth muscle cell migration. Microvessel density and leucocyte infiltration were increased in Hem compared with NH plaques. Plaque vasculature appeared vulnerable as indicated by the absence of alpha-actin-positive mural cells in most plaque vessels. Despite increased numbers of angiogenic factor-expressing microvessels and leucocytes in Hem plaques, lower levels of vascular endothelial growth factor, placental growth factor, and angiopoietin-1 were found in conditioned media from Hem plaques. However, NH and Hem plaques released similar levels of the vascular destabilizing factor, angiopoietin-2. Addition of recombinant angiogenic factors to plaque extracts showed that all factors but angiopoietin-2 were selectively degraded by plasmin and/or elastase released from Hem plaques. Furthermore, conditioned media from Hem plaques showed a reduced ability to induce smooth muscle cell migration.


CONCLUSIONS
Our results provide evidence that immaturity of plaque vessels is associated with the degradation of angiogenic factors by haemorrhage-conveyed leucocytes and proteases.
Le Dall
Julien
J
Inserm and Univ. Paris 7, Cardiovascular Haematology, Bio-Engineering and Remodelling, Bichat-Claude Bernard Hospital, France.
Ho-Tin-Noé
Benoît
B
Louedec
Liliane
L
Meilhac
Olivier
O
Roncal
Carmen
C
Carmeliet
Peter
P
Germain
Stéphane
S
Michel
Jean-Baptiste
JB
Houard
Xavier
X
D000199:Actins / Q000032:analysis; D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D042683:Angiopoietin-1 / Q000378:metabolism; D050197:Atherosclerosis / Q000378:metabolism*; D002465:Cell Movement; D005341:Fibrinolysin / Q000502:physiology; D006470:Hemorrhage / Q000378:metabolism*; D006801:Humans; D019272:Leukocyte Elastase / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D055806:Microvessels / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D018919:Neovascularization, Physiologic; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
C467630:ANGPT1 protein, human; D000199:Actins; D043925:Angiogenesis Inducing Agents; D042683:Angiopoietin-1; D008565:Membrane Proteins; C098069:PIGF protein, human; D042461:Vascular Endothelial Growth Factor A; D019272:Leukocyte Elastase; D005341:Fibrinolysin