Rho kinase-1 mediates cardiac fibrosis by regulating fibroblast precursor cell differentiation
Rho kinase-1 mediates cardiac fibrosis by regulating fibroblast precursor cell differentiation
复制标题
DOI：
10.1093/cvr/cvp135
复制
发表时间：
2009-08-01
影响因子：
10.8
通讯作者：
Entman, Mark L.
中科院分区：
文献类型：
--
作者：
Haudek, Sandra B.;Gupta, Damon;Entman, Mark L.
来源于单核细胞、血源性前体细胞的高度增殖的CD 34（+）/CD 45（+）成纤维细胞在小鼠缺血/再灌注心肌病（I/RC）模型中纤维化的发展中起关键作用。人单核细胞在体外经单核细胞趋化蛋白1（MCP-1）诱导的跨内皮迁移（TEM）后分化为成纤维细胞。由于Rho相关激酶-1（ROCK-1）与纤维化和白细胞TEM有关，我们研究了其在I/RC中的作用。我们发现ROCK-1（-/-）小鼠没有出现I/RC所特有的纤维化和心功能障碍：与野生型相比，ROCK-1（-/-）小鼠心脏显示I/RC诱导的α-平滑肌肌动蛋白（+）成纤维细胞和CD 34（+）/CD 45（+）成纤维细胞前体的数量明显减少。从接受I/RC的ROCK-1（-/-）小鼠分离的心脏成纤维细胞较大且增殖缓慢，与从假手术处理的心脏分离的成纤维细胞相似。我们还进行了体外试验，其中人外周血单核细胞（PBMC）迁移通过内皮细胞响应MCP-1。在迁移之前，将PBMC与ROCK-1靶向小干扰RNA孵育以沉默ROCK-1表达。我们发现，ROCK-1蛋白减少80%并不抑制TEM，但显著减少单核细胞分化成成纤维细胞的数量> 20倍。我们的数据暗示了ROCK-1在分化中的重要作用，但在成熟为心脏成纤维细胞的单核细胞的TEM中不起作用。这些细胞介导非适应性纤维化。
Highly proliferative, CD34(+)/CD45(+) fibroblasts derived from monocytic, blood-borne precursor cells play a critical role in the development of fibrosis in a murine ischaemic/reperfusion cardiomyopathy (I/RC) model. The differentiation of human monocytes into fibroblasts in vitro occurs after transendothelial migration (TEM) induced by monocyte chemoattractant protein 1 (MCP-1). Because Rho-associated kinase-1 (ROCK-1) has been implicated in fibrosis and leukocyte TEM, we investigated its involvement in I/RC.We subjected mice with genetic deletion of ROCK-1 to I/RC. We found that ROCK-1(-/-) mice did not develop the fibrosis and cardiac dysfunction characteristic for I/RC: compared with wild-type, ROCK-1(-/-) hearts showed markedly lower numbers of I/RC-induced alpha-smooth muscle actin(+) fibroblasts and CD34(+)/CD45(+) fibroblast precursors. Isolated cardiac fibroblasts from ROCK-1(-/-) mice undergoing I/RC were large and slowly proliferating, similar to fibroblasts isolated from sham-treated hearts. We also performed in vitro assays in which human peripheral blood mononuclear cells (PBMC) migrated through endothelial cells in response to MCP-1. Prior to migration, PBMC were incubated with ROCK-1-targeting small interfering RNA to silence ROCK-1 expression. We found that an 80% reduction of ROCK-1 protein did not inhibit TEM, but significantly reduced the amount of mononuclear cells that differentiated into fibroblasts by > 20-fold.Our data implicate an important role for ROCK-1 in the differentiation, but not in the TEM of monocytes that mature into cardiac fibroblasts. These cells mediate non-adaptive fibrosis.