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骨形态发生蛋白-2 联合丹酚酸 B 促进大鼠骨髓间充质干细胞向心肌细胞分化。

BMP-2 combined with salvianolic acid B promotes cardiomyocyte differentiation of rat bone marrow mesenchymal stem cells.

机构信息

Department of Histology and Embryology, Hebei North University, Zhangjiakou, China.

Department of Ultrasound, The First Affiliated Hospital of Hebei North University, Zhangjiakou, China.

出版信息

Kaohsiung J Med Sci. 2017 Oct;33(10):477-485. doi: 10.1016/j.kjms.2017.06.006. Epub 2017 Jul 13.

DOI:10.1016/j.kjms.2017.06.006
PMID:28962818
Abstract

The present study tested the hypotheses that bone morphogenetic protein 2(BMP-2) combined with salvianolic acid B(Sal-B) enhance the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) towards cardiomyocytes in vitro. BMSCs were treated with BMP-2 and Sal-B, alone or in combination, for 72 h, then added new media (excluding inductive substance) and cultured for 4 weeks. The morphologic characteristics, surface antigens and mRNA expression of several transcription factors were also assessed. We found that they could all be identified by the positive staining for cardiomyocyte-specific proteins, desmin and cardiac troponin T, in these cells. Furthermore, the mRNA expression of GATA-4 and Nkx2.5 genes was slightly increased on day 7, enhanced on day 14 and decreased on day 28 while α-MHC gene was not expressed on day 7, but expressed slightly on day 14 and enhanced on day 28. The expression of these cardiac-specific markers in treatment groups were all significantly higher than those in the control group, respectively (P < 0.05). Transmission electron microscopy showed that BMSCs in treatment groups all had myofilaments, z line-like substances and mitochondria. Taken together, these results indicate that BMP-2 combined with Sal-B promotes myocardial differentiation of BMSCs, which may represent a potential therapeutic strategy for the treatment of ischemic heart disease.

摘要

本研究旨在验证以下假设

骨形态发生蛋白 2(BMP-2)与丹酚酸 B(Sal-B)联合使用可促进大鼠骨髓间充质干细胞(BMSCs)向心肌细胞分化。将 BMSCs 分别用 BMP-2 和 Sal-B 处理,或联合处理,持续 72 小时,然后加入新的培养基(不含诱导物质)并培养 4 周。还评估了几种转录因子的形态特征、表面抗原和 mRNA 表达。我们发现,这些细胞中的心肌细胞特异性蛋白、结蛋白和心肌肌钙蛋白 T 的阳性染色可识别它们。此外,GATA-4 和 Nkx2.5 基因的 mRNA 表达在第 7 天略有增加,在第 14 天增强,在第 28 天减少,而α-MHC 基因在第 7 天不表达,但在第 14 天略有表达,在第 28 天增强。与对照组相比,治疗组中这些心脏特异性标志物的表达均显著升高(P < 0.05)。透射电镜显示,治疗组中的 BMSCs 均具有肌丝、Z 线样物质和线粒体。综上所述,这些结果表明,BMP-2 联合 Sal-B 可促进 BMSCs 的心肌分化,这可能代表一种治疗缺血性心脏病的潜在治疗策略。

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