Lu Dong-feng, Yao Yan, Su Zi-zhuo, Zeng Zhao-hua, Xing Xiao-wen, He Zhi-yu, Zhang Chunxiang
Department of Cardiology and Sino-US Cardiovascular Research Center, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Department of Pharmacology and Cardiovascular Research Center, Rush Medical College, Rush University, Chicago, Illinois, United States of America.
PLoS One. 2014 Apr 1;9(4):e93222. doi: 10.1371/journal.pone.0093222. eCollection 2014.
Under myocardial microenvironment, bone marrow-derived mesenchymal stem cells (MSCs) can transdifferentiate into cardiomyocytes (CMs). However, the role of histone deacetylase 1 (HDAC1) in this directed differentiation process remains unclear. The current study is to determine the acetylation regulatory mechanisms that may be involved in the directed CM differentiation from MSCs. MSCs isolated from male Sprague-Dawley (SD) rats were marked with Ad-EGFP and co-cultured with CMs. Flow cytometry was used to sort EGFP-positive (EGFP+) MSCs from the co-culture system. Then, the expression of cardiac troponin T (cTnT) in these MSCs was detected by immunofluorescence assay. In addition, HDAC1 levels at different co-culture times were measured by quantitative real-time polymerase chain reaction (QT-PCR) and Western blotting. At 4 days after co-culture with CMs, the MSCs began to expression detectable levels of cTnT. The expression of HDAC1 in CMs was much lower than that in MSCs. After co-culture with CMs, the expression of HDAC1 in MSCs was significantly decreased in a time dependent manner. In addition, our recent study has also identified that knockdown of the HDAC1 could promote the directed differentiation of MSCs into CMs. The results suggest that HDAC1 has a negative correlation with cardiac cell differentiation from MSCs under a myocardial microenvironment. HDAC1 might play an important role in the directed differentiation of MSCs into CMs in heart.
在心肌微环境下，骨髓间充质干细胞（MSCs）可转分化为心肌细胞（CMs）。然而，组蛋白去乙酰化酶1（HDAC1）在这一直接分化过程中的作用仍不清楚。当前研究旨在确定可能参与MSCs向CMs直接分化的乙酰化调控机制。从雄性Sprague-Dawley（SD）大鼠分离的MSCs用Ad-EGFP标记，并与CMs共培养。采用流式细胞术从共培养体系中筛选出EGFP阳性（EGFP+）的MSCs。然后，通过免疫荧光分析检测这些MSCs中心肌肌钙蛋白T（cTnT）的表达。此外，通过定量实时聚合酶链反应（QT-PCR）和蛋白质免疫印迹法检测不同共培养时间下HDAC1的水平。与CMs共培养4天后，MSCs开始表达可检测水平的cTnT。CMs中HDAC1的表达远低于MSCs。与CMs共培养后，MSCs中HDAC1的表达呈时间依赖性显著下降。此外，我们最近的研究还发现，敲低HDAC1可促进MSCs向CMs的直接分化。结果表明，在心肌微环境下，HDAC1与MSCs向心肌细胞的分化呈负相关。HDAC1可能在心脏中MSCs向CMs的直接分化中起重要作用。
