基于Wnt信号途径探讨肌源性干细胞向造血方向分化的调控机制及当归补血汤的干预作用

Muscle derived stem cells (MDSCs), one of pluripotent stem cells, was shown to be able to differentiated into hematopoietic cells under specific circumstances so it is a line of very promising seed cells to restore hematopoietic function in transplantation recipient. Our previous study (No：30772881) has proved that Danggui Buxue Decoction (DBD), a representative herb that tonifys qi and generates blood, can significantly improve hematopoietic microenvironment and restore hematopoietic function in recipient. But its mechanism is to be elucidated. Wnt signaling was reported to involve in regulating stem cell differentiation, through modifying multiplefactors in hematopoietic process. Therefore, we propose that improvement of hematopoietic microenvironment by DBD through Wnt signal could promote MDSCs to differentiate into blood cells. We intend to explore the mechanism of the MDSCs differentiation under intervention with DBD, to enrich the connotation of tonifying qi and generating blood using DBD and to expound the important scientific significance of MDSCs as seed cells in transplant medicine through co-culturing MDSCs and BMSCs , molecular biology , gene chip and RNAi both in vivo and in vitro.

研究证实：具有多向分化潜能的肌源性干细胞（MDSCs）在特定微环境中可分化为造血细胞,在移植治疗中,将成为重建造血的最有希望的种子细胞来源。前期（No.30772881）提示:补气生血方药当归补血汤有明显的改善造血微环境促MDSCs移植受体造血重建的能力，然其深入机制尚待明晰。近期研究发现Wnt信号是调控干细胞分化的关键，有可能从多环节影响造血过程，故在临床和前期研究确切的基础上，提出当归补血汤干预造血微环境，通过Wnt信号调控可促MDSCs血向分化。拟通过在体和离体实验，建立MDSCs/BMSCs共培养模型，及分子生物学技术，以基因芯片与RNAi相结合，重点研究造血微环境下Wnt信号通路调控MDSCs造血分化的作用机制，从中医整体观角度，找到可进行有效干预的新靶点，探讨当归补血汤促MDSCs造血分化的分子机制，该研究对丰富补气生血方药的内涵，及移植治疗中的种子细胞筛选有着重要的科学价值。

研究证实：具有多向分化潜能的肌源性干细胞（MDSCs）在特定微环境中可分化为造血细胞,在移植治疗中,将成为重建造血的最有希望的种子细胞来源。前期（No.30772881）提示:补气生血方药当归补血汤有明显的改善造血微环境促MDSCs移植受体造血重建的能力，然其深入机制尚待明晰。本课题研究发现Wnt信号是调控干细胞分化的关键，从多环节影响造血过程，故在临床和前期研究确切的基础上，提出当归补血汤干预造血微环境，通过Wnt信号调控可促MDSCs血向分化。通过在体实验，当归补血汤给药后，尾静脉移植原代分离出的雄性转基因鼠MDSCs，观察受体小鼠成活状况；在离体实验部分，建立了MDSCs/BMSCs共培养模型，分别收集不同的细胞培养液进行造血集落培养试验及造血重建试验；将基因芯片与RNAi相结合，并通过分子生物学技术检测Wnt信号通路上关键靶点Wnt3a、β-catenin、Wnt10b及其下游靶基因CyclinD1、c-Myc等的表达水平，同时应用统计学方法分析其相关性，从Wnt信号通路调控入手，观察信号通路上造血相关基因Wnt10b、Wnt3、Wnt3a等的表达，明晰外源性MDSCs移植受体促造血恢复，重点研究造血微环境下Wnt信号通路调控MDSCs造血分化的作用机制，从中医整体观角度，找到了可进行有效干预的新靶点，探讨当归补血汤促MDSCs造血分化的分子机制，该研究对丰富补气生血方药的内涵，及移植治疗中的种子细胞筛选有着重要的科学价值。