摘要
目的 观察骨髓间充质细胞（bone marrow mesenchymal stem cells,BMSCs）上清液对缺氧缺血性脑损伤（Hypoxic ischemic brain damage,HIBD）新生大鼠海马胶质纤维酸性蛋白（Glial fibrillary acidic protein,GFAP）表达及学习记忆能力的影响,探讨BMSCs移植治疗HIBD大鼠的机制.方法 将7日龄新生SD大鼠随机分成假手术组、模型组、细胞组和上清液组,每组20只.采用结扎左侧颈总动脉、术后持续缺氧2h建立HIBD模型.模型组大鼠于造模术后1周经左侧脑室注入0.01 moL/L的PBS溶液2μl,细胞组和上清液组大鼠于术后1周分别以相同方法注入等量的BMSCs和BMSCs上清液;假手术组仅分离左颈总动脉,但不结扎,术后不进行缺氧干预,其余处理同模型组.术后8周Morris水迷宫试验检测大鼠的学习记忆功能,免疫组化检测大鼠海马齿状回GFAP的表达.结果 模型组探查训练时在目标象限的时间（Tl）和对位探查训练时在目标象限的时间（T2）分别为（15.40±2.80）s、（16.45± 1.16）s,均较假手术组缩短[T1（19.96±2.57）s、T2（25.32±2.54）s,P＜0.05];细胞组[T1（17.54±1.80）s、T2（18.99±1.47）s]和上清液组[T1（17.40±2.37）s和T2（17.96±1.09）s]分别均较模型组延长（均P＜0.05）,但细胞组和上清液组T1和T2比较差异无统计学意义（P＞0.05）.模型组海马齿状回GFAP阳性物质积分光密度（Integral optical density,IOD）值（15.26± 1.49）较假手术组（12.82±2.56）增加（P＜0.05）,细胞组和上清液组IOD值分别为：（18.13±1.97）、（17.38±1.64）,均较模型组增加（P＜0.05）,但细胞组和上清液组IOD值之间差异无统计学意义（P＞0.05）.结论 BMSC上清液经侧脑室注入能够改善HIBD大鼠的学习记忆能力,主要机制可能为MSCs分泌的某些细胞因子促进了HIBD大鼠的神经修复.
Objective To analyze the expression of glial fibrillary acidic protein（GFAP） in hippocampal of rats with hypoxic-ischemic brain damage（HIBD） after injection of bone marrow stromal cells（BMSCs） supernatant into the lateral ventricle of the neonate rats,assess the ability of learning and memory,and explore the mechanism.Methods 7-day-old SD rats were randomly divided into sham operation group,model group,cells and supernatant group,with 20 rats in each group.The HIBD model was established via the ligation of left carotid arteries followed by 2-hour hypoxia.One week later,rats in control group were injected with 0.01 mol/L PBS 2 μl via the left ventricle.Meanwhile,the cell and the supernatant groups were injected with BMSCs and supernatant harvested from BMSC culture,respectively,via the same route.In the sham operation group,the left carotid arteries were separated but not ligated,and no hypoxia treatment was imposed on this group.They also received 0.01 mol/L PBS injection 2 μl per weak post surgery.8 weeks later,Morris water maze test was performed to assess the learning and memory,and the expression of GFAP in the dentate gyrus of rats was measured by immunohistochemistry.Results Morris water maze showed that the searching time of the probe trail（T1：（15.40±2.80） s） and reversal probe trail （T2：（16.45± 1.16） s） of the model group was shorter than that of the sham operation group （T1：（19.96±2.57） s,T2：（2：25.32±2.54）s,P〈0.05）,while the searching time of the cell group（T1：（17.54± 1.80）s,T2：（18.99± 1.47） s） and supernatant group （T1：（17.40±2.37） s,T2：（17.96± 1.09） s） was prolonged compared with that of the model group （P〈0.05）.No significant difference between the cell group and the supernatant group （P〉0.05）.The integral optical density （IOD） value of GFAP positive cells was higher in the model group than that in the sham operation group（15.26±1.49 vs 12.82±2.56,P〈0.05）,while the IOD of cell group（18.13±1.97） and that of the supernatant group（17.38± 1.64） were higher than that of the model group （P〈0.05）.However,there was no significant difference between the cell group and the supernatant group （9〉0.05）.Conclusion BMSC supernatant injected through lateral ventricle can improve the HIBD rats＆#39; abilities of learning and memory.The mechanism might be that MSCs secrete some cytokines to promote central nervous system repair.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2015年第11期982-985,共4页
Chinese Journal of Behavioral Medicine and Brain Science
基金
四川省泸州市科技局课题[2008-S-12（4/4）]
关键词
骨髓间充质细胞
上清液
缺氧缺血性脑损伤
胶质纤维酸性蛋白
记忆
Bone marrow mesenchymal stem cells
Supernatant
Hypoxic isehemic brain damage
Glial fibrillary acidic protein
Memory
