Revuelta-López Elena, Soler-Botija Carol, Nasarre Laura, Benitez-Amaro Aleyda, de Gonzalo-Calvo David, Bayes-Genis Antoni, Llorente-Cortés Vicenta
Cardiovascular Research Center, CSIC-ICCC, IIB Sant Pau, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain.
ICREC (Heart Failure and Cardiac Regeneration) Research Program, Health Sciences Research Institute Germans Tries i Pujol, Badalona (Barcelona), Spain.
J Cell Mol Med. 2017 Sep;21(9):1915-1928. doi: 10.1111/jcmm.13113. Epub 2017 Apr 4.
Left ventricular (LV) remodelling after myocardial infarction (MI) is a crucial determinant of the clinical course of heart failure. Matrix metalloproteinase (MMP) activation is strongly associated with LV remodelling after MI. Elucidation of plasma membrane receptors related to the activation of specific MMPs is fundamental for treating adverse cardiac remodelling after MI. The aim of current investigation was to explore the potential association between the low-density lipoprotein receptor-related protein 1 (LRP1) and MMP-9 and MMP-2 spatiotemporal expression after MI. Real-time PCR and Western blot analyses showed that LRP1 mRNA and protein expression levels, respectively, were significantly increased in peri-infarct and infarct zones at 10 and 21 days after MI. Confocal microscopy demonstrated high colocalization between LRP1 and the fibroblast marker vimentin, indicating that LRP1 is mostly expressed by cardiac fibroblasts in peri-infarct and infarct areas. LRP1 also colocalized with proline-rich tyrosine kinase 2 (pPyk2) and MMP-9 in cardiac fibroblasts in ischaemic areas at 10 and 21 days after MI. Cell culture experiments revealed that hypoxia increases LRP1, pPyk2 protein levels and MMP-9 activity in fibroblasts, without significant changes in MMP-2 activity. MMP-9 activation by hypoxia requires LRP1 and Pyk2 phosphorylation in fibroblasts. Collectively, our in vivo and in vitro data support a major role of cardiac fibroblast LRP1 levels on MMP-9 up-regulation associated with ventricular remodelling after MI.
心肌梗死后左心室重构是心力衰竭临床病程的关键决定因素。基质金属蛋白酶(MMP)激活与心肌梗死后左心室重构密切相关。阐明与特定MMP激活相关的质膜受体是治疗心肌梗死后不良心脏重构的基础。本研究的目的是探讨低密度脂蛋白受体相关蛋白1(LRP1)与心肌梗死后MMP-9和MMP-2时空表达之间的潜在关联。实时聚合酶链反应(PCR)和蛋白质印迹分析表明,在心肌梗死后10天和21天,梗死周边区和梗死区的LRP1信使核糖核酸(mRNA)和蛋白表达水平均显著升高。共聚焦显微镜显示LRP1与成纤维细胞标志物波形蛋白高度共定位,表明LRP1主要由梗死周边区和梗死区的心脏成纤维细胞表达。在心肌梗死后10天和21天,LRP1在缺血区心脏成纤维细胞中也与富含脯氨酸酪氨酸激酶2(pPyk2)和MMP-9共定位。细胞培养实验表明,缺氧可增加成纤维细胞中LRP1、pPyk2蛋白水平和MMP-9活性,而MMP-2活性无显著变化。缺氧诱导的MMP-9激活需要成纤维细胞中LRP1和Pyk2磷酸化。总之,我们的体内和体外数据支持心脏成纤维细胞LRP1水平在心肌梗死后与心室重构相关的MMP-9上调中起主要作用。