Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells
Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells
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DOI：
10.1016/j.ejcb.2007.09.003
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发表时间：
2008-03-01
影响因子：
6.6
通讯作者：
Simionescu, Maya
中科院分区：
文献类型：
--
作者：
Burlacu, Alexandrina;Rosca, Ana-Maria;Simionescu, Maya
骨髓基质细胞(BMSC)可以分化为包括心肌细胞在内的多种细胞类型，在心肌梗死的细胞治疗中具有一定的应用价值。为了增加骨髓间充质干细胞的生肌性，我们考察了去甲基化试剂5-氮胞苷诱导的转化程度。取成年大鼠胫骨分离骨髓间充质干细胞，接种后1d，加入5MU的5-氮胞苷培养2 4h。每周重复一次治疗，并评估肌肉特异性蛋白质和基因的表达。结果表明，培养的细胞失去了骨钙素和碱性磷酸酶的天然表达，并开始表达连接蛋白43。暴露于5-氮胞苷的骨髓间充质干细胞在14天时诱导出一种类似肌细胞的表型，包括肌肉特异性蛋白(肌小节α-肌动蛋白、肌钙蛋白T、结蛋白、α-肌动蛋白和GATA-4)和基因(GATA-4、myoD)、结蛋白和α-肌动蛋白的表达，大量线粒体和肌丝；但后者没有形成肌节。虽然一些生肌标记物也出现在未经处理的细胞中，但暴露于5-氮胞苷后，钙通道的反应增强，结蛋白和肌动蛋白D基因表达增加三倍，α-肌动蛋白基因和蛋白表达增加两倍。综上所述，5-氮胞苷对体外培养的骨髓间充质干细胞中肌肉特异性蛋白和基因的表达有促进作用。值得注意的是，肌源性分化发生在很短的时间内。启动间充质细胞向心肌细胞分化可能在改善心肌缺血后肌肉损失的细胞方法中有重要的应用。(C)2007年爱思唯尔股份有限公司。版权所有。
Bone marrow stromal cells (BMSC) can differentiate into various cell types including myocytes, which may be valuable in cellular therapy of myocardial infarction. In an attempt to increase the myogenic commitment of BMSC, we investigated the extent of conversion induced by the demethylation agent 5-azacytidine. BMSC isolated from the adult rat tibia were exposed in culture to 5 mu M 5-azacytidine for 24 h, 1 day after seeding. The treatment was repeated at weekly intervals and the expression of muscle-specific proteins and genes was assessed. The results revealed that cultured cells lost the native expression of osteocalcin and alkaline phosphatase as a function of time and began to express connexin 43. Exposure to 5-azacytidine of BMSC induced, at 14 days, a myocyte-resembling phenotype that included the expression of muscle-specific proteins (sarcomeric alpha-actin, troponin T, desmin, alpha-actinin, and GATA-4) and genes (GATA-4, myoD), desmin, and alpha-actinin), numerous mitochondria and myofilaments; however, the latter did not form sarcomeres. Although some of these myogenic markers also appeared in untreated cells, exposure to 5-azacytidine induced an enhanced response of calcium channels, as well as a threefold increase in desmin and myoD gene expression and a twofold increase in a-actinin gene and protein expression above the control values. In conclusion, the results demonstrate a promoting effect of 5-azacytidine on the expression of muscle-specific proteins and genes in BMSC in culture. Notably, the myogenic differentiation takes place over a short period of time. Priming of mesenchymal cells to cardiomyogenic differentiation may have significant applications in cellular approaches to ameliorate muscle loss after myocardial ischemia. (C) 2007 Elsevier GmbH. All rights reserved.