Villarreal F J, Lee A A, Dillmann W H, Giordano F J
Department of Medicine, University of California, San Diego 92103, USA.
J Mol Cell Cardiol. 1996 Apr;28(4):735-42. doi: 10.1006/jmcc.1996.0068.
Transforming growth factor-beta 1 (TGF-beta 1) is known to regulate cardiac cell function and its overexpression in the heart is thought to contribute to the development of cardiac hypertrophy and fibrosis. We wished to develop a high efficiency gene transfer method that could be used both in vitro and in vivo and result in the overexpression of TGF-beta 1. For this purpose, we constructed a replication-deficient human adenovirus 5 vector encoding for human TGF-beta 1 and used for control purposes an adenovirus lacZ vector. The adenovirus 5 construct was capable of infecting neonatal rat cardiac myocytes, fibroblasts and VSMCs. Of the three cell types, cardiac myocytes appear more susceptible to infection by the adenovirus 5 construct as assessed through beta-galactosidase staining. Infection of cardiac fibroblasts, myocytes and VSMCs with the hTGF-beta 1 adenovirus leads to the expression of hTGF-beta 1 mRNA and enhanced levels of bioactive and total TGF-beta 1 protein. Infection with hTGF-beta 1 adenovirus also results in enhanced levels of collagen type III gene expression in VSMCs and fibroblasts whereas in cardiac myocytes it leads to increased levels for sarcomeric and beta-actin. Thus, this adenoviral vector might be used for the exploration of in vivo effects of altered levels of cardiac TGF-beta 1.
已知转化生长因子β1（TGF-β1）可调节心脏细胞功能，其在心脏中的过度表达被认为与心脏肥大和纤维化的发展有关。我们希望开发一种高效的基因转移方法，该方法可在体外和体内使用，并导致TGF-β1的过度表达。为此，我们构建了一种编码人TGF-β1的复制缺陷型人腺病毒5载体，并使用腺病毒lacZ载体作为对照。腺病毒5构建体能够感染新生大鼠心肌细胞、成纤维细胞和平滑肌细胞。在这三种细胞类型中，通过β-半乳糖苷酶染色评估，心肌细胞似乎更容易被腺病毒5构建体感染。用hTGF-β1腺病毒感染心脏成纤维细胞、心肌细胞和平滑肌细胞会导致hTGF-β1 mRNA的表达以及生物活性和总TGF-β1蛋白水平的提高。用hTGF-β1腺病毒感染还会导致平滑肌细胞和成纤维细胞中III型胶原基因表达水平的提高，而在心肌细胞中则会导致肌节和β-肌动蛋白水平的增加。因此，这种腺病毒载体可用于探索心脏TGF-β1水平改变的体内效应。
