基于Wnt信号通路探讨益气养阴中药纳米微球对骨髓间质干细胞诱导神经细胞表型稳定的研究

Spinal cord injury and neurodegenerative diseases were common in clinical.The central nervous system after injury was very difficult to recover due to the mammalian neuron lack of the ability to regenerate，and it was difficult to achieve satisfactory clinical results yet. The clinical research showed that Shenmai injection had good protecting effect on nerve cells.Based on this，the project constructed Chinese Medicine nanospheres with Shenmai injection,Chitosan and Sodium alginate.We induced rat bone marrow stem cells (MSCs) to differentiate into neurons through slow releasing of the Shenmai injection and growth factor bFGF auxiliarily. We investigated the mechanism of the Chinese medicine nanospheres which has the function of Supplementing Qi and nourishing Yin in maintaining the induced neuron cell phenotype stable by observation the expression of the neuron cell BaX/Bcl2， neurotransmitter after induction ,and also the influence on Wnt Signaling pathway ,etc. we hoped that we could harvest more neuron cells with good function and ensure the long-term efficacy and stability of the neural defects reparation with tissue engineering neuron cells. This research established on the induced neuron cells declining easily and tried to explore a new method that could maintain the cell phenotype stable. So we could accumulate some experiences in the extant investigation and promote the modernization process of Chinese medicine.

脊髓损伤与神经系统退行性疾病是临床常见病、多发病，但由于成年哺乳动物的神经元缺乏再生能力，中枢神经系统受损后极难恢复，因此临床上治疗该类疾病仍难以取得满意疗效。研究证实参麦液对神经细胞具有良好的保护作用，在此基础上，本课题以参麦液、壳聚糖、海藻酸钠构建中药复合体纳米微球，通过对参麦液的缓控释，辅助生长因子bFGF定向诱导大鼠骨髓基质干细胞（MSCs）分化为神经元细胞，对比观察诱导后神经元细胞Bax／Bcl-2等神经递质的表达以及Wnt信号通路的影响，探讨中药纳米微球在定向调控MSCs分化神经元细胞、维持诱导后神经元样细胞表型、促进神经元递质表达并抑制其凋亡等方面的作用机理，保证组织工程化神经元细胞移植修复神经缺损的长期有效性和稳定性。本研究以诱导后神经细胞易衰退老化为切入点，寻找维持细胞表型稳定的新方法，可望在中医药研究的深度与广度上积累一定的经验，同时也促进中医药现代化的进程。

脊髓损伤与神经系统退行性疾病是临床常见病、多发病，但由于成年哺乳动物的神经元缺乏再生能力，中枢神经系统受损后极难恢复，因此临床上治疗该类疾病仍难以取得满意疗效。研究证实参麦液对神经细胞具有良好的保护作用，在此基础上，本课题以中药、壳聚糖、海藻酸钠构建中药复合体微球，通过对中药红参的缓控释，辅助生长因子 bFGF定向诱导人骨髓基质干细胞（MSCs）分化为神经元细胞，对比观察诱导后神经元细胞 Bax／Bcl-2 等神经递质的表达以及 Wnt 信号通路的影响，探讨中药微球在定向调控 MSCs分化神经元细胞、维持诱导后神经元样细胞表型、促进神经元递质表达并抑制其凋亡等方面的作用机理，保证组织工程化神经元细胞移植修复神经缺损的长期有效性和稳定性。结果显示中药微球载药稳定，释药性良好，可通过wnt3a诱导MSCs细胞向神经元分化，可抑制细胞凋亡，对诱导后神经细胞表型稳定具有一定作用，从而保证组织工程化神经元细胞移植修复神经缺损的长期有效性。本研究以诱导后神经细胞易衰退老化为切入点，寻找维持细胞表型稳定的新方法，可望在中医药研究的深度与广度上积累一定的经验。