Perez-Ilzarbe Maitane, Agbulut Onnik, Pelacho Beatriz, Ciorba Cristina, San Jose-Eneriz Edurne, Desnos Michel, Hagège Albert A, Aranda Pablo, Andreu Enrique J, Menasché Philippe, Prósper Felipe
Hematology, Cardiology and Cell Therapy, Clínica Universitaria and Division of Cancer, Foundation for Applied Medical Research, Division of Cancer, University of Navarra, Pamplona, Spain.
Eur J Heart Fail. 2008 Nov;10(11):1065-72. doi: 10.1016/j.ejheart.2008.08.002. Epub 2008 Sep 20.
The discrepancy between the functional improvements yielded experimentally by skeletal myoblasts (SM) transplanted in infarcted myocardium and the paucity of their long-term engraftment has raised the hypothesis of cell-mediated paracrine mechanisms.
We analyzed gene expression and growth factors released by undifferentiated human SM (CD56(+)), myotubes (SM cultured until confluence) and fibroblasts-like cells (CD56(-)). Gene expression revealed up-regulation of pro-angiogenic (PGF), anti-apoptotics (BAG-1, BCL-2), heart development (TNNT2, TNNC1) and extracellular matrix remodelling (MMP-2, MMP-7) genes in SM. In line with the gene expression profile, the analysis of culture supernatants of SM by ELISA identified the release of growth factors involved in angiogenesis (VEGF, PIGF, angiogenin, angiopoietin, HGF and PDGF-BB) as well as proteases involved in matrix remodelling (MMP2, MMP9 and MMP10) and their inhibitors (TIMPs). Culture of smooth muscle cells (SMC), cardiomyocytes (HL-1) and human umbilical vein endothelial cells (HUVECs) with SM-released conditioned media demonstrated an increased proliferation of HUVEC, SMC and cardiomyocytes (p<0.05) and a decrease in apoptosis of cardiomyocytes (p<0.05). Analysis of nude rats transplanted with human SM demonstrated expression of human-specific MMP-2, TNNI3, CNN3, PGF, TNNT2, PAX7, TGF-beta, and IGF-1 1 month after transplant.
Our data support the paracrine hypothesis whereby myoblast-secreted factors may contribute to the beneficial effects of myogenic cell transplantation in infarcted myocardium.
梗死心肌中移植的骨骼肌成肌细胞（SM）在实验中产生的功能改善与其长期植入的缺乏之间的差异，引发了细胞介导的旁分泌机制的假说。
我们分析了未分化的人SM（CD56(+)）、肌管（培养至汇合的SM）和成纤维细胞样细胞（CD56(-)）释放的基因表达和生长因子。基因表达显示，SM中促血管生成（PGF）、抗凋亡（BAG-1、BCL-2）、心脏发育（TNNT2、TNNC1）和细胞外基质重塑（MMP-2、MMP-7）基因上调。与基因表达谱一致，通过ELISA对SM培养上清液的分析确定了参与血管生成的生长因子（VEGF、PIGF、血管生成素、血管生成素、HGF和PDGF-BB）以及参与基质重塑的蛋白酶（MMP2、MMP9和MMP10）及其抑制剂（TIMPs）的释放。用SM释放的条件培养基培养平滑肌细胞（SMC）、心肌细胞（HL-1）和人脐静脉内皮细胞（HUVEC），结果显示HUVEC、SMC和心肌细胞的增殖增加（p<0.05），心肌细胞凋亡减少（p<0.05）。对移植了人SM的裸鼠的分析表明，移植后1个月，人特异性MMP-2、TNNI3、CNN3、PGF、TNNT2、PAX7、TGF-β和IGF-1表达。
我们的数据支持旁分泌假说，即成肌细胞分泌的因子可能有助于梗死心肌中肌源性细胞移植的有益效果。
