EGb761脑脊液药理学方法介导循环式干细胞移植治疗痴呆研究

With the arrival of aging , the economic and social problems caused by dementia become highlights. Looking for a new , effective , simple operation , small side effects treatment is a urgent need for doctors. Our previous date have indicated that Mesenchymal Stem Cells(MSCs) using cerebrospinal fluid induced can produce a sufficient number of Neural Stem Cells(NSCs) , which transplanted through Intrathecal injection can be helpful to improve the neurological functions, the quality of life and the ability of living of dementia patients. But between different cerebrospinal fluid mesenchymal stem cells the induction mechanism not entirely consistent. What’s more,we found that Ginkgo biloba extract761(EGb761) improved the cognitive impairment by reducing the damages of neurons in hippocampus, enhancing the SOD, GSH-px activities, the ratio of Bcl-2 to Bax and decreasing the level of MDA,the expression of Caspsae3. Based on the above date,we made the assumption that Ginkgo biloba extract cerebrospinal fluid pharmacology may be a good inducing microenvironment. In the next study,we will use the pharmacology to induce MSCs to NSCs ,and using the Stem Cells to treatment of dementia. The result of the experiment will be proved by using inverted optical microscope and electron microscope, immunohistochemical and fluorescent immunohistochemistry, RT-PCR and flow cytometry analysis in three level ,which including ethology, brain slice electrophysiology and molecular biological.While we also will compare the pros and cons of the stem cells transplantation path.This study may offer new ideas for treatment of dementia.

老龄化使得痴呆带来的问题日益严峻，我们前期研究发现模拟脑内微环境的脑脊液诱导法所获得的干细胞通过鞘内注射移植可以改善痴呆患者的神经功能，但不同脑脊液的诱导作用并不完全一致。同时还发现银杏叶提取物761 可通过提高SOD、GSH活性、降低MDA水平，提高Bcl-2／Bax比值、降低Caspsae3表达减少神经元损伤而表现出对认知的保护作用。基于此，本项目首次利用银杏叶提取物761脑脊液药理学方法，应用倒置光学显微镜及电子显微镜、免疫组化及荧光免疫组化、RT-PCR及流式细胞分析等手段技术从动物行为学、脑片电生理及分子生化3 个水平研究中药脑脊液药理学在诱导干细胞分化中的优势。同时将诱导获得的干细胞再通过脑脊液途径移植到模型动物体内实现“治疗与获取诱导剂”的同步，并在移植前后评估认知相关标志的改变，从而探索出一种循环型的、临床可操作的个体化干细胞移植治疗模式去满足痴呆患者的干细胞治疗需求。

神经元丢失是阿尔茨海默病（Alzheimer's disease，AD）患者功能损伤的根本原因之一，目前对AD的治疗多以缓解症状为主，而不能解决神经元丢失问题。基于此，我们在获得资助后应用免疫组化及荧光免疫组化、RT-PCR及Westernblot、流式细胞分析等方法在细胞和在体水平探讨EGb761脑脊液药理学的建立及其在AD干细胞治疗中的应用，发现：1）给6月龄AD小鼠（5×FAD）长期EGb761（20, 30mg/kg）腹腔注射后能够明显抑制由于环境应激诱导的小胶质细胞长期激活而诱发的海马椎体神经元损伤、增加海马脑区新生神经元的数量、促进新生神经元分化后整合入原有学习记忆网络实现突触网络结构的重建，最终提高其学习记忆能力；2）获得的“EGb-CSF”（即使来源AD）在促进干细胞的存活、增殖和分化及抑制神经炎症等方面的作用比单独的EGb761强；3）“EGb761- BMSCs”联合使用可能是较好的神经再生、神经保护方案，这为将来EGb761在干细胞移植治疗中的应用提供了另一种思路，即配合BMSCs移植，做BMSCs的“卫士”。但以上具体机制均不清楚，因此值得进一步深入探讨和研究。