转VEGF基因内皮祖细胞移植治疗阿尔茨海默病模型小鼠的研究
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Research of Alzheimer disease model mouse implanted with endothelial progenitor cells transfected with VEGF gene
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    摘要:

    目的 将体外诱导的血管内皮祖细胞(EPCs)转入血管内皮生长因子(VEGF)基因,并定 向移植到AD 模型鼠脑内海马区,观察该部位及邻近部位的血管重建以及内皮细胞功能的改善情况,为 AD的治疗提供研究基础。方法 采用体外分离培养小鼠骨髓源性EPCs。构建的携带人VEGF165 基 因的pcDNA3.1 质粒转染至EPCs,观察EPCs 表达VEGF 情况。采用APP/PS1 双转基因鼠模型,将转染人 VEGF165 基因的EPCs 移植至模型鼠脑内,观察小鼠行为学的变化、Aβ 含量,以及血管的形成情况。结 果 将转染VEGF基因的EPCs移植入小鼠的海马区后,动物在第4,5,6 天进行空间导航试验发现,与 对照组比较,平均潜伏期明显缩短(P< 0.05);免疫组化结果显示,与对照组、EPCs 组比较,EPCs+VEGF 组额叶皮层和海马区Aβ含量明显减少(P< 0.01),海马区CD34 阳性细胞数更高(P < 0.01)。结论 将 EPCs+VEGF 移植入鼠脑内,可以发现小鼠的记忆能力增强,原因可能与促进新生血管的生成、Aβ 清除 有关。

    Abstract:

    Objective Vascular endothelial progenitor cells( EPCs) induced in vitro were transferred into vascular endothelial growth factor( VEGF) gene and transplanted into the hippocampus of Alzheimer disease (AD) model rats. To observe the reconstruction of blood vessels and the improvement of endothelial cell function in this area and adjacent areas, so as to provide basis for the treatment of AD. Methods This experiment used EPCs obtained from the bone marrow of mouse and kept in in vitro cultures. By constructing pcDNA3.1 plasmids carrying human VEGF165 gene and transfecting the EPCs, how the gene affects EPCs and its expression were observed. Using the APP/PS1 double transgenic model rats, human VEGF165 gene transfected EPCs was implanted into model rat's brain, to observe rat's behavioral differences, Aβ protein amounts, and blood vessel formation. Results After implanting EPCs transfected with VEGF gene into the hippocampus of mice, the animals were tested on 4, 5 and 6 d. The average latency was significantly shorter than that in the control group (P < 0.05). The results of immunohistochemistry showed that compared with the control group and the EPCs group, the content of Aβ in the frontal cortex and hippocampus of EPCs+VEGF group decreased significantly, and the number of CD34 positive cells in hippocampus were higher( P< 0.01). Conclusions By implanting EPCs+VEGF into brains of mice, the memory ability of mice can be found to be enhanced, which may be related to the promotion of angiogenesis and the clearance of Aβ protein.

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王玉琳 刘晶 张朝东.转VEGF基因内皮祖细胞移植治疗阿尔茨海默病模型小鼠的研究[J].神经疾病与精神卫生,2018,18(2):
DOI :10.3969/j. issn.1009-6574.2018.02.011.

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  • 在线发布日期: 2018-05-16