小鼠脑室内注射神经干细胞裂解液促进谷氨酸盐诱导的兴奋性神经元损伤的修复  

Repair Following Glutamate-induced Excitotoxic Neuronal Damage Mediated by Intracerebroventricular Injection of Cell-free Filtrate of Neural Stem Cell Lysates in Adult Mice

作　　者：于立坚[1,2] 马娟[1] 马润娣[1,2] 张永平[1] 房娟芝[1] 张霄瑜[1,3,2] 于廷曦[1,4,2] 

机构地区：[1]广东海洋大学海洋药物重点实验室,湛江524025 [2]佛山德溥生物医药科技有限公司,佛山528231 [3]Department of Otorhinolaryngology-Head and Neck Surgery,University of Maryland School of Medicine,md21228 [4]Cell Biology Group,Department of Surgery,Department of Pathology,University of Maryland School of Medicine and Baltimore Veterans Affairs Medical Center,md21201

出　　处：《中国细胞生物学学报》2011年第10期1086-1093,共8页Chinese Journal of Cell Biology

基　　金：广东海洋大学研究生基金资助项目~~

摘　　要：先前的研究已经证实,阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液具有改善抑郁症样模型大鼠的行为学障碍、上调其海马和大脑皮质神经生长因子(nerve growth factor,NGF)和脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的表达、增加海马神经干细胞(neural stem cells,NSCs)/神经前体细胞(neural progenitor cells)增殖的效果。该研究的目的在于探讨神经干细胞裂解液的无细胞滤液(cell-free filtrate ofneural stem cell lysates,FNSCL)脑室内注射促进谷氨酸盐诱导的成年小鼠兴奋性神经元损伤修复的可能性。成年小鼠谷氨酸单钠(monosodiumglutamate,MSG,2.0g/(kg·d))灌胃,连续10日,造成兴奋性神经元损伤模型。自孕15 d的昆明种小鼠取胎脑,分离、培养神经干细胞,免疫细胞化学法检测巢蛋白(nestin)抗原,制备神经干细胞裂解液的无细胞滤液。MSG+NSCs组动物在MSG灌胃后接收脑室内NSCs移植,MSG+FNSCL组动物在MSG灌胃后接受脑室内FNSCL注射。Y-迷宫分辨学习试验检测神经功能恢复情况;组织病理学方法检查鼠脑形态结构变化。结果显示,无论是神经干细胞裂解液无细胞滤液脑室内注射还是神经干细胞脑室内移植都同样能促进谷氨酸盐诱导的成年小鼠兴奋性毒性神经元损伤的修复。这一发现提示,不仅神经干细胞,而且神经干细胞裂解液的无细胞滤液也可以用于促进脑损伤的修复。Our previous study demonstrated that cell-free filtrate of sodium ferulate-induced and differentioned PC 12 cell lysates significantly attenuated chronic mild stress-induced depression-like behavioural disorders, up-regulated hippocampal and cerebral cortex expressions of nerve growth factor （NGF） and brain-derived neurotrophic factor （BDNF）, and increased hippocampal number of neural stem cells （NSC）/neural progenitor cells in mice. The present study was undertaken to investigate the possibility of the repair following glutamate （monosodium glutamate, MSG）-induced excitotoxic brain injury mediated by intracerebroventricular injection of cell-free filtrate of neural stem cell lysates （FNSCL） in adult mice. Mouse NSCs were isolated from the brains of embryos at 15 day postcoitum （dpc）. The expression of nestin, a special antigen for NSC, was detected by immunocytochemistry. Cellfree filtrate of NSCs was prepared from the NSC lysates. The animals in the MSG group received intragastric （ig） administration of MSG （2.0 g/（kg.d） for 10 days）, the animals in the MSG＋NSCs group received intracerebroventricular transplantation of NSCs （approximately 1.0× 10^5 cells）, and the animals in the MSG＋FNSCL group received intracerebroventricular injection of 10 uL of FNSCL （approximately 1.0× 10^5 cells） separately on day 1 and day 10 after 10-d MSG exposure. The mice in control and MSG groups received intracerebroventricular injection of Dulbecco＇s modified Eagle＇s medium （DMEM） instead of NSCs or FNSCL. On 12 day after the last intracerebroventricular injection of FNSCL or transplantation of NSCs, Y-maze test was performed, and then the histopathology of animal brains was studied to analyze MSG-induced functional and morphological changes and the effects of intracerebroventricular injection of FNSCL and transplantation of NSCs on the repair of MSG-induced excitotoxic brain injury. The results showed that both intracerebroventricular injection of FNSCL and intracerebroven

关 键 词：脑修复 神经干细胞裂解液 神经干细胞因子 神经干细胞 兴奋性毒性脑损伤 小鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]