Objective To investigate the effect of mannitol combined with the vein graft supported with bone marrow mesenchymal stem cell ( BMSC ) on the cognitive function and the expression of brain-derived neurotrophic factor ( BDNF ) in brain tissue of vascular dementia ( VD ) rats.Methods The BMSCs of rat were isolated and purified by whole bone marrow culture and adherence screening methods.Seventy nine healthy male SD rats were randomly assigned to sham operation group ( n = 10 ) and model ( n = 69) groups.A VD model was established by the ligation of the bilateral common carotid arteries at an interval of 3 days.Four weeks after modeling, the 32 rats in the model group were randomly allocated into mannitol pretreatment BMSC transplantation group( tail intravenous injection of mannitol 1.5 g/kg, tail intravenous injection of 1 × l06 mL BMSCs 1 mL after pretreatment for 1 to 30 minutes), BMSC transplantation group( injecting the same amount of BMSCs but not mannitol), and medium control group( injecting the same amount of basal medium).After removing the dead rats, there were 9, 11, and 8 rats, respectively in the above 3 groups.Seven rats were included in the sham operation group.Morris water maze test was used to detect cognitive function of the rats, and an enzyme-linked immunosorbent assay was used to detect the BDNF content in brain tissue 4 weeks after intervention.Results ①Comparing the 4 groups at the same time points, the escape latency in the BMSC transplantation group was shorter than that in the medium control group ( all P ＜ 0.01 ), and residence time in the platform quadrant was prolonged ( P ＜ 0.05).The BMSC transplantation group after mannitol pretreatment was also significantly shorter than the escape latency in the BMSC transplantation group (P ＜ 0.01 ), the residence time in the platform quadrant was prolonged ( P ＜ 0.05 ) , and they were all close to the level of the sham operation group ( P ＞ 0.05 ).②The BDNF content in the BMSC transplantation group was significantly higher than that in the medium control group in the frontal cortical and hippocampal regions (P ＜ 0.01 ).The mannitol pretreatment BMSC transplantation group was more significantly higher than the BMSC group (P ＜ 0.01 ) , however, they were lower than the level in the sham operation group (P ＜ 0.01 ).Conclusion Compared with intravenous injection of BMSCs alone, the improvement of cognitive function of the VD rat model treated with mannitol combined with BMSC transplantation is much better; the BDNF expression in cortex and hippocampus is higher.%目的 探讨甘露醇预处理后骨髓间充质干细胞(BMSCs)静脉移植对血管性痴呆(VD)大鼠认知功能及脑组织中脑源性神经生长因子(BDNF)表达量的影响.方法 全骨髓培养结合细胞贴壁法分离纯化大鼠BMSCs.取健康雄性sD大鼠79只,随机分为假手术组(10只)和模型组(69只).采用间隔3d分别结扎双侧颈总动脉法制备VD模型.造模后4周,将纳入的32只模型组大鼠随机分为甘露醇预处理BMSCs移植组(简称甘露醇预处理组;尾静脉注射甘露醇1.5 g/kg,预处理10～30min后,经尾静脉注射1X106/ml BMSCs 1 m1)、BMSCs移植组(注射等量的BMSCs,不注射甘露醇)和培养基对照组(注射等量的基础培养液).剔除死亡大鼠,上述3组分别有9、11、8只,假手术组7只纳入统计.干预后4周,采用Morriss水迷宫实验检测大鼠认知功能,酶联免疫吸附法检测大鼠脑组织中BDNF的含量.结果 ①4组相同时间点的比较,BMSCs移植组比培养基对照组逃避潜伏期缩短(均P<0.01),平台象限滞留时间延长(P<0.05);甘露醇预处理组比BMSCs移植组逃避潜伏期明显缩短(P<0.01),平台象限滞留时间延长(P<0.05),且均与假手术组水平接近(P>0.05).②在额叶皮质和海马区,BMSCs移植组的BDNF含量均高于培养基对照组(P<0.01);甘露醇预处理组则高于BMSCs组(P<0.01),但均低于假手术组的水平(P<0.01).结论 与单独静脉注射BMSCs比较,甘露醇预处理后BMSCs静脉移植治疗VD大鼠模型,认知功能改善程度更佳;额叶皮质和海马中BDNF的表达量更高.
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