甘露醇对骨髓间充质干细胞治疗血管性痴呆大鼠行为学及海马CA3区突触素表达水平的影响

The effects of bone marrow mesenchymal stromal cells transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3 region in hippocampus of vascular dementia rats

目的探讨甘露醇预处理对骨髓间充质干细胞（bonemaiTowmesenchymalstromalcells，BMSCs）静脉移植治疗血管性痴呆（vasculalr dementia，VD）模型大鼠行为学及海马CA，区突触素表达的影响。方法以全骨髓贴壁法培养大鼠BMSCs。采用间隔3d双侧颈总动脉永久性结扎法制备VD模型，设立假手术组。造模4周后将VD大鼠按随机数字表法分为模型组、培养基组、甘露醇组、BMSCs组、甘露醇预处理BMSCs组，分别给予相应的实验干预措施。观察干预4周后实验大鼠的行为学表现及应用免疫组织化学法检测海马CA，区突触素的表达水平。结果甘露醇预处理BMSCs组行为学表现较其他实验组有明显改善，其第5天逃避潜伏期（s）比BMSCs组、模型组、培养基组、甘露醇组明显缩短（9．3±2．9，14．1±3．5，23．5±4．4，22．8±4．4，23．2±2．8，F=43．900，P=0．ooo），平台象限滞留时间（S）比BMSCs组、模型组、培养基组、甘露醇组明显延长（40．8±6．3，34．9±5．8，26．4±4．8，27．4±7．0，28．5±6．2，F=13．000，P=0．000），其海马CA3区突触素表达水平（39624±7798）亦比BMSCs组、模型组、培养基组、甘露醇组明显提高（27060±4668，18294±6446，19956±4244，18946±4953，F=39．206，P=0．000）。结论甘露醇预处理后静脉移植BMSCs显著改善VD大鼠行为学表现，并使其海马CA，区突触素表达增加。甘露醇预处理明显提高静脉移植BMSCs治疗VD大鼠的效果。

Objective To investigate the effects of bone marrow mesenchymal stromal cells （BMSCs） transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3 region in hippocampus of vascular dementia （VD） rats. Methods The BMSCs of rats were isolated and purified by the whole bone marrow adherence method. The rats were subjected for permanent ligation of bilateral common carotid arteries at an interval of 3 days for each carotid artery. At the same time, Sham group was set in parallel. Four weeks after modeling, the VD rats were divided randomly into five groups： （1） VD control group; （2） culture media group; （3） mannitol group; （4） BMSCs group; （5） mannitol with BMSCs group. Morris water maze performance and synaptophysin expression in the CA3 region in hippocampus were observed at 4 weeks after transplantation. Results The morris water maze performance significantly improved in mannitol with BMSCs group when compared with BMSCs group, VD control group, culture media group, mannitol group. Moreover, the escape latency of fifth day decreased significantly （（9.3±2.9）, （14.1 ±3.5）, （23.5±4.4）, （22.8±4.4）, （23.2±2.8） s, F=43.900, P = 0. 000） ）, and the platform quadrant residence time increased significantly （ （ 40. 8 ±6. 3 ）, （ 34. 9 ±5.8）, （26.4±4.8）, （27.4±7.0）, （28.5±6.2） s, F=13.000, P=0.000））. The synaptophysin expressions of the hippocampal CA3 region were significantly increased in the mannitol with BMSCs group （39 624 ±7798 ） when compared with BMSCs group, VD control group, culture media group, mannitol group （27 060 ±4668, 18 294 ±6446, 19 956±4244, 18 946 ±4953, F=39.206, P=0.000）. Conclusions Intravenous BMSCs transplantation after mannitol pretreatment improves the behavioral performance of VD rats and facilitates the synaptophysin expression of hippoeampal CA3 region in VD rats than BMSCs transplantation alone. Mannitol pretreatment can amplify the therapeutic effect of intravenous BMSCs transplantation in VD rats.