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目的探讨内皮祖细胞(EPCs)移植对动脉粥样硬化(AS)模型大鼠脑缺血再灌注损伤(IRI)后学习记忆能力与脑顶叶皮质结构的影响。方法高脂膳食饲养建立30只动脉粥样硬化大鼠模型,随机分为AS组,IRI组和EPCs移植组。采集骨髓分离EPCs并体外扩增培养,检测其表面标记物的表达;第7天采用线栓法制作局灶性IRI模型,建模成功后1d EPCs移植组经尾静脉移植EPCs,IRI组与AS组给予等量体积的磷酸盐缓冲液。移植后7d检测各组大鼠的行为能力、脑组织血管内皮生长因子(VEGF)含量及其mRNA表达与其结构的病理改变。结果培养24h后见细胞贴壁生长逐渐变为梭形;第3天细胞明显增殖集落形成;第5天细胞集落逐渐增大呈现克隆样生长;第7天细胞汇合达80%;第10～14天细胞基本铺满瓶底呈铺路石样密集排列。荧光显微镜下,DIL-ac-LDL和FITC-UEA-1双荧光染色的细胞数占贴壁细胞数的75%以上。与IRI组相比,EPCs移植后大鼠的学习记忆能力较IRI组明显改善,VEGF含量及其mRNA表达显著下降(P<0.05)。光镜下,EPCs移植组大鼠脑缺血侧顶叶皮质Caspase-3和胶质细胞原纤维酸性蛋白(GFAP)阳性神经元均较IRI组明显下降(P<0.05)。结论 EPCs移植能改善AS模型大鼠脑IRI后的学习记忆能力、减轻脑组织的病理损害,这些变化提示EPCs促进了神经的修复。
Objective To investigate the effects of endothelial progenitor cells (EPCs) transplantation on the learning and memory ability and the parietal cortex structure after cerebral ischemia-reperfusion injury (IRI) in atherosclerotic rat models. Methods A total of 30 rat models of atherosclerosis were established by high fat diet and were randomly divided into AS group, IRI group and EPCs transplantation group. The EPCs were collected and expanded in vitro to detect the expression of their surface markers. On the 7th day, the focal IRI models were made by the suture method. EPCs were transplanted into the EPCs transplanted via the tail veins 1 d after successful modeling. Groups were given an equal volume of phosphate buffer. At 7 days after transplantation, the behavioral abilities, the content of vascular endothelial growth factor (VEGF) and the mRNA expression of the brain tissue and the pathological changes of the tissues in each group were detected. Results After 24 hours of culture, the adherent cells grew into spindle shape gradually. On the third day, the cells proliferated significantly. On the fifth day, the colonies grew gradually and showed clonal growth. On the seventh day, the cells reached 80% Basal cells covered the bottom of the paving stone-like arrangement. Under the fluorescence microscope, DIL-ac-LDL and FITC-UEA-1 double-stained cells accounted for more than 75% of adherent cells. Compared with IRI group, the learning and memory abilities of rats after EPCs transplantation were significantly improved compared with those of IRI group, and the levels of VEGF and mRNA were significantly decreased (P <0.05). Under light microscope, Caspase-3 and glial fibrillary acidic protein (GFAP) positive neurons in the ischemic parietal cortex of EPCs transplantation group were significantly decreased compared with IRI group (P <0.05). Conclusion EPCs transplantation can improve the learning and memory abilities of IR rats after cerebral IRI and alleviate the pathological damage of brain tissue. These changes suggest that EPCs promote nerve repair.