丹参诱导胎盘间充质干细胞体外定向分化为心肌细胞的有效成分及相关机制

Transplantation of stem cells in treating ischemic heart disease has not been widely applied in clinic because of its certain limitations, mainly reflected in: in vivo we cannot accurately control the differentiation of stem cells into cardiomyocytes, in vitro we are lack of safe and effective myocardial differentiation inducing agents. Our previous study indicated that hPDMSCs were easier to get in comparison with BMSCs and could be induced to differentiate into cardiomyocytes without relying on cytotoxic agents or co-culture technique but with relying on dan-shen root. Based on our previous work, we intend to use dan-shen root and its effective monomer composition (salvianolic acid B, danshensu, protocatechuic aldehyde, tanshinone II A, tanshinone I, cryptotanshinone) to induce human placenta-derived mesenchymal stem cells (hPDMSCs) to differentiate into cardiomyocytes and use canonical and noncanonical Wnt signal pathway inhibitors to intervent differentiation system. By detecting the expession changes of cardiac specific transcription factors and characteristics of genes and proteins and Wnt signal pathway related genes and proteins, we want to make clear about the effective components and mechanism about dan-shen root on cardiomyogenic differentiation of hPDMSCs. The results of the study can provide a good inducer agent using for cardiomyogenic differentiation. It also has an important practical significance in the traditional Chinese medicine intervention in myocardial tissue engineering.

如何控制干细胞定向分化为心肌细胞一直是干细胞研究领域的热点和难点，寻找安全、高效的心肌细胞分化诱导剂一直是心肌组织工程领域研究学者的梦想。我们前期研究发现，丹参能够诱导胎盘间充质干细胞体外定向分化为心肌细胞。本项目在前期工作的基础上，拟用丹参及其单体成分：丹酚酸B、丹参素、原儿茶醛、丹参酮ⅡA、丹参酮Ⅰ、隐丹参酮诱导胎盘间充干细胞向心肌细胞分化，并用经典和非经典的Wnt 信号通路抑制剂进行干预，通过检测干预前、后不同分化阶段心肌特异性转录因子和特征性基因及蛋白、Wnt 信号通路中相关基因和蛋白的表达变化情况，明确丹参诱导hPDMSCs体外定向分化为心肌细胞的有效成分及其相关机制。本项目的研究结果能为心肌组织工程中研发安全、高效的心肌分化诱导剂提供有价值的理论和实验依据，对我国传统中药介入心肌组织工程具有重要的示范意义。

如何控制干细胞定向分化为心肌细胞一直是干细胞研究领域的热点和难点，寻找安全、高效的心肌细胞分化诱导剂一直是心肌组织工程领域研究学者的梦想。在前期发现丹参能够诱导人胎盘间充质干细胞（human placenta-derived mesenchymal stem cells，hPDMSCs）体外定向分化为心肌细胞且效果优于目前常用的共培养诱导法的基础上，通过无毒剂量的丹参及其有效的单体成分丹参素、丹酚酸B、原儿茶醛、丹参酮Ⅰ、丹参酮ⅡA、隐丹参酮干预hPDMSCs，检测干预前、后细胞增殖速率、细胞形态学变化、心肌特征性基因和蛋白的表达变化情况及Wnt信号通路相关基因和蛋白的表达变化情况，从而明确了丹参诱导hPDMSCs体外定向分化为心肌细胞的有效成分为丹参酮ⅡA, 且诱导效率明显高于丹参，Wnt/β-catenin信号通路在丹参酮ⅡA诱导hPDMSCs心肌样分化中发挥着重要作用。本项目的研究结果能为心肌组织工程中研发安全、高效的心肌分化诱导剂提供有价值的理论和实验依据，对我国传统中药介入心肌组织工程具有重要的示范意义。