骨髓基质细胞移植改善APP/PS1转基因小鼠的认知功能

Bone marrow stromal cells grafting improves the cognitive disorders of APP/PS1 transgenic mice

目的探究骨髓基质细胞(Bone marrow stromal cells,BMSCs)移植治疗对APP/PS转基因的阿尔兹海默病(Alzheimer's disease,AD)模型小鼠认知功能障碍的改善作用。方法分离培养C57BL/6小鼠的BMSCs,诱导其分化为神经细胞,通过相差显微镜技术和免疫细胞化学荧光染色技术鉴定其表型与分化。通过显微注射技术,将绿色荧光蛋白(Enhanced green fluorescent protein,eGFP)转基因小鼠(生后3个月)来源的BMSCs注入APP/PS转基因小鼠(生后12个月)侧脑室内后,以荧光显微镜技术检测移植细胞的存活和整合情况,HE染色检测AD模型小鼠海马区萎缩的恢复程度并以Morris水迷宫试验与旷场试验检测移植治疗后小鼠认知功能障碍的改善情况。结果分离自C57BL/6小鼠的BMSCs在增殖培养基中贴壁生长,形态呈纺锤形,免疫表型CD271+/CD45-++,经神经分化诱导培养可形成β-tubulin-III神经元和GFAP星形胶质细胞。GFP转基因小鼠来源的BMSCs移植入APP/PS1转基因小鼠脑内可存活并与宿主海马区神经组织有效整合。移植治疗3周后宿主小鼠海马区面积显著增加(<0.001),水迷宫实验逃生潜伏期明显缩短(<0.01),旷场试验评分显著提高(<0.05)。结论骨髓基质细胞移植治疗可有效改善APP/PS1转基因的AD模型小鼠的认知功能障碍。

Objective To investigate the improving effects of bone marrow stromal cells（BMSCs） grafting therapy on the cognitive disorders of APP/PS1 transgenic Alzheimer＇s disease（AD） model mice. Methods BMSCs were isolated from C57BL/6 mice（3 months postnatal, P3 mon）, cultured and differentiated into neural cells. The phenotype and differentiation of these BMSCs were identified by phase contrast microscopy and immunocytochemistry. BMSCs derived from enhanced green fluorescent protein（eGFP） transgenic mice were injected into the lateral ventricle of APP/PS1 transgenic mice （12 months postnatal/P12 mon） by microinjection. The survival and integration of grafted BMSCs were observed by microscopy, HE staining used to assess the restoration of the atrophy of hippocampus area of AD model mice, and Morris＇s water maze and open field test were used to evaluate the improvement of cognitive disorders after BMSCs transplantation. Results C57BL/6 mice derived BMSCs showed adherent growth in proliferation medium, spindle shape and CD271＋/CD45- positive or [3 -tubulin-lIF positive or GFAP＋ positive. The eGFP transgenic mice derived BMSCs survived and integrated into hippocampus of AD mice, the average size of hippocampus was significantly increased（P〈0.001）, the average escaping latency reduced evidently（P〈0.01） and the score by open field test was increased notably（P 〈0.05）, compared to AD model mice. Conclusion BMSCs grafting therapy can improve the cognitive disorders of APP/PS l transgenic AD model mouse effectively.