神经干细胞移植对老年痴呆模型鼠基底前脑小白蛋白阳性神经元和学习记忆能力的影响  

Effect of neural stem cells transplantation on parvalbumin-positive neurons of the basal forebrain and abilities of learning and memory in a rat model of senile dementia

作　　者：谷海刚[1] 龙大宏[2] 李晓滨[1] 张贵平[3] 苏涛[4] 李佳楣[2] 冷水龙[2] 

机构地区：[1]广州医学院组织胚胎学教研室,广东省广州市510182 [2]广州医学院解剖学教研室,广东省广州市510182 [3]广州医学院药理学教研室,广东省广州市510182 [4]广州医学院神经科学研究所,广州医学院附属第二医院,广东省广州市510260

出　　处：《中国组织工程研究与临床康复》2008年第12期2235-2239,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金(06022674)~~

摘　　要：目的:基底前脑是老年性痴呆患者脑皮质下神经元丢失最严重的区域,实验拟验证经神经干细胞移植治疗后老年痴呆鼠基底前脑小白蛋白阳性神经元的变化以及对其空间学习记忆能力方面的影响。方法:实验于2005-12/2006-07在广州医学院解剖学教研室完成。①动物:清洁级SD雄性大鼠28只,随机数字表法分为3组:正常对照组8只、模型对照组8只、细胞移植组12只。另取10只新生SD鼠用于神经干细胞的分离培养。实验过程中对动物的处置符合动物伦理学标准。②实验方法:模型对照组、细胞移植组大鼠切断左侧穹隆海马伞,建立老年性痴呆动物模型。利用无血清培养技术获得鼠基底前脑神经干细胞,吸取2.5×1010L-1细胞悬液4μL,术后即刻细胞移植组损伤侧行细胞移植,坐标为前囟+0.6mm,外侧+0.6mm插入,腹侧-5.5mm。③实验评估:移植后4周,采用Y型迷宫对各组动物进行学习记忆能力检测。麻醉后取脑制备组织切片,ABC法免疫组化染色结合图像分析观察各组大鼠基底前脑小白蛋白阳性神经元的变化。结果:①对空间学习记忆能力的影响:细胞移植组空间学习能力、记忆能力均显著高于模型对照组(P<0.05或P<0.01),基本达到正常水平(P>0.05)。②对小白蛋白阳性神经元的影响:与正常对照组比较,模型对照组损伤侧内侧隔核和斜角带核的小白蛋白阳性神经元数目分别减少62.5%和30.4%;细胞移植组降低幅度明显小于模型对照组(P<0.01)。与正常对照组比较,模型对照组损伤侧内侧隔核、斜角带核小白蛋白阳性神经元的面积、周长、灰度均显著降低(P<0.05或0.01);细胞移植组上述指标较模型对照组均显著增加(P<0.05或0.01)。③相关性分析:小白蛋白阳性神经元数目与大鼠在Y型迷宫中的学习次数呈显著负相关(r=-0.76^-0.79,P<0.01),与记忆能力呈显著正相关(r=0.78~0.84,P<0.01)。结论:神经干细胞移植对老年性�AIM： The loss of neurons is the severest in the basal forebrain of senile dementia patients. This study was aimed to verify the effects of neural stem cells （NSC） transplantation on the parvalbumin （PV）-positive neurons in the basal forebrain and the abilities of leaning and memory in the rat model of senile dementia. METHODS： This study was done in the Department of Anatomy, Guangzhou Medical College from December 2005 to July 2006. （1）Animals： SPF class adult male SD rats （n=28） were randomly divided into three groups： normal control group （n =8）, model control group （n =8）, and NSC group （n =12）. Furthermore, new-born SD rats （n =10） were used to isolate and culture NSC. All the manipulations were in accordance with the animal ethnic standard.（2）Methods： The rats of model group and NSC group were made by left fimbria-fornix transaction. NSCs were obtained from the basal forebrain by using the serum-free culturing. And 4 μL NSC suspension （2.5×10^10 L^-1） was stereotaxically implanted into the injured lesion （coordinates： anterior-posterior, ＋0.6 mm; lateral, ＋0.6 mm; dorsal-ventral, -5.5 mm from bregma） instantly after fimbria-fornix transaction.（3）Evaluation： Four weeks postoperation, the learning and memory abilities were measured by Y-maze test. Then, the brains were taken out after anesthesia and cut on a microtome. The number of PV-positive neurons in the basal forebrain was analyzed by using ABC immunohistochemical staining method and image analysis. RESULTS： （1）Effects on learning and memory abilities： The abilities in NSC group were significantly higher than that in model control group （P 〈 0.05 or P 〈 0.01）, and almost reached the normal level （P 〉 0.05）.（2）Effects on PV-positive neurons： Compared with normal control group, PV-positive neurons in the medial septum （MS） and the vertical limb of the diagonal band of Broca （VDB） in model control group decreased by 62.5% and 30.4%, respectively. The dec

关 键 词：神经干细胞 老年性痴呆 小白蛋白阳性神经元 学习 记忆 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]