甘露醇对骨髓基质细胞移植治疗缺血性脑损伤的研究

The Study of Mannital's Effect on the Treatment of MCAO by Transplanting Bone Marrow Stromal Cells

目的利用甘露醇的开放血脑屏障及细胞保护作用,探索其增加静脉移植的骨髓基质细胞(BMSCs)进入脑内数量的可行性,为下一步BMSCs或载基因细胞静脉移植应用于临床奠定理论基础。方法利用贴壁筛选法分离、培养、扩增BMSCs至第5代,移植前经BrdU标记。线栓法制作大脑中动脉缺血模型(MCAO),通过尾静脉进行注射移植。将动物模型分为三组,空白对照组:注射甘露醇;常规治疗组:移植BMSCs;治疗组:移植BMSCs后注射甘露醇。在第1、7、14、21天分别行NSS评分并取脑组织制作切片,在荧光显微镜下记数BrdU阳性细胞,以确定BMSCs的分布及存活情况。结果BMSCs能较早期进入脑内存活,且集中于缺血灶周围。在同时间点,应用甘露醇移植组大鼠的NSS评分较其它两组差别有统计学意义。结论甘露醇通过其开放血脑屏障及细胞保护的作用,能增加进入脑内并存活的BMSCs的数量。

Objective By use of Mannital＇ s liberalizing BBB and protecting ceils, to search for the feasibility of raising the number of BMSCs that migrate into injured rat brains, and to provide the further theoretical foundation for the clinical application of the intravenous transplantation of bone marrow stromal cells or carrying- gene cells. Methods We used the method of adherence screening to isolate BMSCs from rat bone marrow, then cultured BMSCs and proliferated to the 5th generation, marked with BrdU and transplanted them to MCAO models through 14th and 21th day, NSS scores were measured the tail vein. Three groups were made. At the 1st , 7th, and brain tissue from different rats was extracted and then made into slices. By observing the number of BrdU positive cells under fluorescence - microscope, we determined the location and survival situation of BMSCs. Results By means of the effect of Mannital＇ s liberalizing BBB and protecting cells, BMSCs can more easily migrate into injured brains to live, while in normal tissues they are hardly available. The differences of NSS scores among these three groups were striking. Conclusions By means of its effect of liberalizing BBB and protecting cells, Mannital facilitates the migration of BMSCs＇ into ischemic brain tissue and its survival.