Coutard Michèle, Touat Ziad, Houard Xavier, Leclercq Anne, Michel Jean-Baptiste
INSERM, U 698, and University Paris 7, FR-75877 Paris, France.
J Vasc Res. 2010;47(4):355-66. doi: 10.1159/000265569. Epub 2009 Dec 16.
BACKGROUND/AIM: The intraluminal thrombus (ILT) is considered to participate in abdominal aortic aneurysm (AAA) evolution. To assess whether this role proceeds via ILT influence on biological activity of the AAA wall, we studied the relationships between the levels of some relevant proteases and microparticles (MP) released by ILT versus wall in rat experimental AAAs.
Two weeks after elastase perfusion, ILT and AAA wall were incubated in cell culture medium and studies were performed on conditioned media. As shown by gelatin zymography, ILT released higher amounts of MMP9 than the wall, whereas the level of MMP2 activation (active/pro) was similar. Levels of elastase and urokinase plasminogen activator, plasmin and MPs, determined, respectively, by casein zymography, substrate hydrolysis and flow cytometry, were higher in ILT than in wall. Aneurysm diameter positively correlated with wall MMP9 levels, MMP2 activation, plasmin activity and MP release. Moreover, wall and ILT levels of pro- and active forms of MMP2, elastase and plasmin were positively correlated. Wall levels of MMP2 activation and plasmin activity also correlated with ILT weight.
The present data suggest that, in this experimental model, ILT may contribute to AAA evolution via its influence on the level of aneurysmal wall protease activity.
背景/目的:腔内血栓(ILT)被认为参与腹主动脉瘤(AAA)的发展。为了评估这种作用是否通过ILT对AAA壁生物活性的影响来实现,我们研究了大鼠实验性AAA中ILT与壁释放的一些相关蛋白酶和微粒(MP)水平之间的关系。
弹性蛋白酶灌注两周后,将ILT和AAA壁在细胞培养基中孵育,并对条件培养基进行研究。如明胶酶谱所示,ILT释放的MMP9量高于壁,而MMP2激活水平(活性/前体)相似。分别通过酪蛋白酶谱、底物水解和流式细胞术测定的弹性蛋白酶、尿激酶型纤溶酶原激活剂、纤溶酶和MPs水平,ILT高于壁。动脉瘤直径与壁MMP9水平、MMP2激活、纤溶酶活性和MP释放呈正相关。此外,MMP2、弹性蛋白酶和纤溶酶的前体和活性形式在壁和ILT中的水平呈正相关。壁MMP2激活水平和纤溶酶活性也与ILT重量相关。
目前的数据表明,在这个实验模型中,ILT可能通过影响动脉瘤壁蛋白酶活性水平而促进AAA的发展。