动物实验和小规模人体临床试验已证实，人脐带间充质干细胞（HUCMSCs ）移植治疗缺血性卒中是安全有效的，然而评价HUCMSCs不同途径移植后在动物活体内存活、分布、迁移、增殖及分化的差异及机制研究明显滞后于功能改善观察和机理的研究，成为干细胞治疗机制探讨的瓶颈因素。我们前期研究表明通心络对大鼠脑缺血后内源性NSCs的增殖与定向分化有明显的促进作用，有助于脑梗死患者移植HUCMSCs后功能改善。在此基础上运用活体生物光学分子成像技术示踪比较大鼠脑缺血后经静脉、动脉、脑室、脑实质内、腰穿移植HUCMSCs后不同时间点大鼠体内HUCMSCs分布、存活、迁移情况的差异及通心络干预后的影响,并在各时间点行组织学检测，与活体示踪法相结合，明确移植入的 HUCMSCs增殖、分化、迁移的机制，实时、动态观察通心络促进脑缺血损伤后HUCMSCs移植的疗效及机制，探讨中医药在干细胞移植治疗脑缺血中的作用。

A lot of animal experiments and some human clinical trials have proved that the transplant of HUCMSCs to treat the ischemic stroke is effective and safe. However, the evaluation of the study about tracting the distribution, survive, migration and differentiation of the stem cells in the living body is obviously less advanced than the observational study and the study of the mechanism. At the same time, it becomes the bottleneck and hotspot of the study about the stem cells. The early studies have showed that tongxinluo capsule can accelerate the proliferation and oriented differentiation. Based on that theory, if we can use the In vivo optical molencular imaging technology tracer technique to compare the distribution, survive, migration and differentiation of the HUCBMCs after the cerebral ischemia of the mice with HUCMSCs transplant through the veins, arteries, ventricles，parenchymal and LP，and the influence of the tonxinluo capsule and do the test in the different timepoints combining the vivo tracer tenique ,then maybe we can figure out the mechanism of the proliferation, differentiation and migration of the HUCBMCs transplant after the cerebral ischemia. It is really meaningful to observe the effect and mechanism of tongxinluo capsule after the cerebral ischemia thereby to testify the "collateral disease" theory. It is of great importance to study the mechanism of tongxinluo capsule and will do good to the communication of the western medicine and traditional Chinese medicine.

本研究运用Lenti-Fire Luciferase (Luc) -eGFP转染HUCMSCs、生物发光成像、PCR等分子生物学技术,采用线栓法制作小鼠MACO模型，观察了经静脉、动脉、脑室、脑实质内、腰穿五种途径移植HUCMSCs后不同时间点小鼠体内HUCMSCs分布、存活、迁移情况的差异。取得以下成果：1、五种途径移植HUCMSCs治疗小鼠脑缺血再灌注损伤，均可减少小鼠MACO模型的脑梗死体积、改善神经功能症状，其中经动脉移植在7-14天时脑梗死体积的减少优于静脉、脑室、脑实质内、腰穿移植（P<0.05）2、于HUCMSCs移植后第24小时、3、7、14、30天，采用IVIS Spectrum system动态连续检测其生物发光信号的强度变化，结果显示，移植后各时间点各组生物发光信号强度显著优于模型组（P<0.05），其中动脉移植组荧光信号强度均强于其它移植途径组（P<0.05）；经中药通心络干预后，移植的HUCMSCs的存活及迁移能力均高于单纯的HUCMSCs移植组效果（P<0.05），表明通心络对HUCMSCs移植后的存活、迁移具有促进作用3、进行小鼠MCAO模型步态参数变异与基因表达关系的研究，发现在第7天，与假手术组相比，MCAO模型中39个步态参数与8个缺血相关基因中的4个基因（MMP9，NOS1，P2RY12和VWF）的mRNA表达水平显著改变（p <0.05),此外，26个步态参数与4个缺血相关基因显著相关（皮尔森相关系数> 0.7或<-0.7）。