人羊膜间充质干细胞静脉移植治疗转基因APP^+小鼠的机制  

Mechanisms underlying intravenous transplantation of human amniotic mesenchymal stem cells for Alzheimer's disease in transgenic APP^+ mice

作　　者：王成春[1,2] 杨波[2,3] 关方霞[1] 李国栋[4] 周长辉[1,2] 周云帆[1] 胡祥[5,6] 谷晨熙[7] 雷宁静[1] 

机构地区：[1]郑州大学生物工程系,河南省郑州市450001 [2]河南省高等学校临床医学重点学科开放实验室,河南省郑州市450052 [3]郑州大学第一附属医院神经外科,河南省郑州市450052 [4]郑州大学药物实验中心,河南省郑州市450052 [5]江苏省干细胞与生物治疗公共技术服务平台,江苏省泰州市225300 [6]深圳市北科细胞工程研究所,广东省深圳市518000 [7]郑州大学临床医学系,河南省郑州市450001

出　　处：《中国组织工程研究与临床康复》2010年第19期3471-3476,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：郑州大学211三期建设项目"干细胞基础与临床研究";河南省科技厅09科技发展计划"干细胞基础与临床研究"(092102310250);江苏省干细胞与生物治疗公共技术服务平台"干细胞与生物治疗公共技术服务中心"(BM2008146)~~

摘　　要：背景:研究发现APP基因与阿尔茨海默病发病密切相关。课题组前期实验发现人羊膜间充质干细胞静脉移植可以促进阿尔茨海默病APP+转基因小鼠的学习,记忆能力改善。目的:探讨人羊膜间充质干细胞尾静脉移植后,能否归巢到小鼠脑组织中,并分化为相关的中枢神经细胞,治疗阿尔茨海默病。方法:无菌条件下体外分离培养人羊膜间充质干细胞,传至第3代将细胞浓度调整为1×109L-1,经尾静脉注入0.5mL至细胞移植组转APP+基因小鼠体内;对照组经尾静脉注入同体积的生理盐水;正常组转APP-基因小鼠不给予任何干预措施。采用免疫组化方法检测、测定Brdu标记的人第3代羊膜间充质干细胞在小鼠脑组织中的表达,各组小鼠脑组织中GFAP、Nestin和NSE的表达。结果与结论:光镜下观察可见,移植组小鼠脑组织中大部分细胞核被染成蓝色,但有一些细胞核被染成棕色或褐色,Brdu呈阳性。与对照组相比,移植组小鼠脑组织中的GFAP增加了近4倍,表达显著增加,甚至超过正常组的表达(P<0.05);Nestin的表达则显著升高,增加了近10%,但是还比正常组低了近20%(P<0.05)。NSE的表达降低了近1/3,但仍高于正常组(P<0.05)。表明人羊膜间充质干细胞尾静脉移植治疗阿尔茨海默病可能是通过羊膜间充质干细胞归巢到转基因APP+小鼠脑组织中,并分化成中枢神经细胞的途径实现的。BACKGROUND：APP gene is closely associated with the onset of Alzheimer＇ disease.Intravenous transplantation of human amniotic mesenchymal stem cell（AMSCs） can promote the learning and memory improvement in transgenic APP＋ mice with Alzheimer＇ disease.OBJECTIVE：To study whether the AMSCs can transfer into the brain tissue of Alzheimer＇s disease mice,and differentiate into the neural cell,then cure the disease after the human AMSC transplantation via tail venous injection.METHODS：Human AMSCs were isolated in vitro sterilely.At the third passage,0.5 mL single cell suspension at 1×109/L was obtained and transplanted by tail venous pathway in transplantation group mice,Mice in the control group were injected with an equal volume of saline.APP-gene mice in the normal group were left intact.5＇-bromo-2-deoxyuridine（BrdU） labeled third-generation AMSCs expression was detected in mice brain tissue by immunohistochemical method.Glial fibrillary acidic protein（GFAP）,Nestin and neuron specific enolase（NSE） expression was measured in the brain tissue of mice from each group.RESULTS AND CONCLUSION：Under an optical microscope,a majority of nuclei in the brain tissue of mice from transplantation group were stained blue,but some nuclei were stained brown,positive for BrdU.Compared with control group,the expression of GFAP in the brain tissue of transplanted mice was increased about 4 times,even more than in the normal group（P 0.05）.The expression of Nestin in the brain tissue of transplanted mice was increased about 10%,but still lower than the normal group nearly 20%（P 0.05）.NSE expression was decreased by 1/3,but still higher compared with the normal group（P 0.05）.Above-mentioned results have shown that human AMSC transplantation for treating Alzheimer＇s disease takes place by AMSCs homing to APP＋ mouse brain tissue and differentiating into neural cells.

关 键 词：人羊膜间充质干细胞 转基因APP+小鼠 尾静脉移植 胶质纤维蛋白 巢蛋白 神经元特异性烯醇化酶 

分 类 号：R394.2[医药卫生—医学遗传学]