Pretreatment of Rat Bone Marrow Mesenchymal Stem Cells with a Combination of Hypergravity and 5-Azacytidine Enhances Therapeutic Efficacy for Myocardial Infarction
Pretreatment of Rat Bone Marrow Mesenchymal Stem Cells with a Combination of Hypergravity and 5-Azacytidine Enhances Therapeutic Efficacy for Myocardial Infarction
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DOI：
10.1002/btpr.558
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Li, Ying-Hui
中科院分区：
文献类型：
--
作者：
Ling, Shu-Kuan;Wang, Rui;Li, Ying-Hui
背景和目的：未修饰的成人骨髓间充质干细胞（BMSCs）在心肌梗死（MI）后的体内心脏分化和功能作用存在争议。我们以前的研究结果表明，超重力促进BMSCs的心肌分化，因此我们推测，离体预处理的BMSCs使用超重力和5-氮杂胞苷（5-Aza）将导致心肌分化，并导致上级的生物学和功能的影响，对心肌梗死心肌的心脏再生。方法：采用结扎大鼠冠状动脉的方法建立心肌梗死模型。分离、培养、扩增大鼠骨髓间充质干细胞，并通过加入超重力（2G）3天和5-Aza（50 lmol/L，24 h）诱导分化为心肌细胞系。大鼠梗死后注射DAPI标记的BMSCs，随机分为5组。A组为对照组，B组为未修饰的BMSCs组，C组为超重力处理组，D组为5-Aza处理组，E组为5-Aza+超重力处理组（n = 6）。结果：RT-PCR检测显示，经超重力和5-Aza处理后，BMSCs表达早期心肌和肌肉标志物加塔-4、MEF-2和Nkx 2 -5。我们还发现超重力可以通过促进HDAC 5的核输出来增强MEF-2的活性。冰冻切片显示，植入的DAPI标记的BMSCs存活，并在植入部位鉴定出血管生成。在B、C、D和E组大鼠中，预处理的BMSC与α-辅肌动蛋白共定位，并且E组大鼠显示出左心室功能的显著更大增加。结论：在移植前用超重力和5-Aza对成人BMSCs进行生物学离体心肌分化是可行的，并且似乎改善了它们在体内的心脏分化以及在大鼠梗死心肌模型中的功能恢复。(C)2011年美国化学工程师学会生物技术。程序，27：473-482，2011
Background and Purpose: The in vivo cardiac differentiation and functional effects of unmodified adult bone marrow mesenchymal stem cells (BMSCs) after myocardial infarction (MI) is controversial. Our previous results suggested that hypergravity promoted the cardiomyogenic differentiation of BMSCs, and thus we postulated that ex vivo pretreatment of BMSCs using hypergravity and 5-azacytidine (5-Aza) would lead to cardiomyogenic differentiation and result in superior biological and functional effects on cardiac regeneration of infarcted myocardium. Methods: We used a rat MI model generated by ligation of the coronary artery. Homogeneous rat BMSCs were isolated, culture expanded, and differentiated into a cardiac lineage by adding hypergravity (2G) for 3 days and 5-Aza (50 lmol/L, 24 h). Rats underwent BMSCs (labeled with DAPI) injection after the infarction and were randomized into five groups. Group A rats received the control medium, Group B rats received unmodified BMSCs, Group C rats received BMSCs treated with hypergravity, Group D rats received BMSCs treated with 5-Aza, and Group E rats received BMSCs treated with 5-Aza and hypergravity (n = 6). Results: After hypergravity and 5-Aza treatment, BMSCs showed positive for the early muscle and cardiac markers GATA-4, MEF-2, and Nkx2-5 with RTPCR. We also found that hypergravity could enhance the activities of MEF-2 via promoting the nuclear export of HDAC5. The frozen section showed that the implanted BMSCs labeled with DAPI survived and angiogenesis was identified at the implantation site. In Groups B, C, D, and E rats, pre-treated BMSCs colocalized with alpha-actinin, and Group E rats showed a significantly larger increase in left ventricular function. Conclusions: The biological ex vivo cardiomyogenic differentiation of adult BMSCs with hypergravity and 5-Aza prior to their transplantation is feasible and appears to improve their in vivo cardiac differentiation as well as the functional recovery in a rat model of the infarcted myocardium. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 473-482, 2011