摘要
背景:目前面神经损伤后的修复主要集中在外周神经干,但面神经损伤后会导致部分中枢运动神经元凋亡。现阶段关于干细胞植入面神经损伤大鼠脑后对面神经核团内凋亡神经元的影响相关报道甚少。目的:观察大鼠面神经损伤后,脑内移植绿色荧光蛋白转基因胎鼠的神经干细胞的成活和迁移情况。设计、时间及地点:随机对照动物实验,于2008-07/12在昆明医学院神经科学研究所完成。材料:孕14～16d的绿色荧光蛋白转基因蛋白小鼠1只,用于制备转基因神经干细胞。清洁级SD雄性大鼠24只,随机分为3组:面神经损伤转基因细胞组12只、面神经损伤细胞培养液组6只、面神经正常转基因细胞组6只。方法:面神经损伤转基因细胞组、面神经损伤细胞培养液组大鼠建立面神经切断模型。造模后1周,行转基因神经干细胞立体定向移植,注射点为前囟后方11.30mm、背侧9.00mm、正中线位置。面神经损伤转基因细胞组、面神经正常转基因细胞组各注入10μL转基因神经干细胞悬液(含5×106个细胞),面神经损伤细胞培养液组注入10μL神经干细胞培养液,4周后制作脑组织冰冻切片。主要观察指标:荧光显微镜观察移植部位绿色荧光蛋白阳性神经干细胞的存活情况,及其向损伤侧面神经核团周围迁移的情况。结果:①移植处:面神经损伤转基因细胞组、面神经正常转基因细胞组均可见数量不等的绿色荧光蛋白阳性细胞,其中部分绿色荧光蛋白阳性细胞位于血管内;面神经损伤细胞培养液组未见绿色荧光蛋白阳性细胞。②面神经核团周围:仅面神经损伤转基因细胞组可见数量不等的绿色荧光蛋白阳性细胞迁移于损伤侧面神经核团周围,而健侧面神经核周围未见绿色荧光蛋白阳性细胞;余2组双侧面神经核周围均未见绿色荧光蛋白阳性细胞。③移植处与面神经核团周围之间:未见绿色荧光蛋白阳性细胞相连。结论:神经干细胞移植入面神经损伤大鼠脑内后,可以向损伤侧面神经核周围迁移。
BACKGROUND： Repair following facial nerve injury mainly focused on peripheral nerve stem, but facial nerve injury may result in apoptosis of central motor neurons. There are few studies concerning effects of stem cell transplantation on apoptotic neurons in rats following facial nerve injury. OBJECTIVE： To explore the survival and migration of transplanted neural stem cells （NSCs） of green fluorescent protein （GFP） transgenic mice in rat brain after facial nerve injury. DESIGN, TIME AND SETTING： The randomized, controlled, animal study was performed at the Neuroscience Institute of Kunming Medical College from July to December 2008. MATERIALS： One GFP transgenic mice pregnant 14-16 days were used for preparing transgenic NSCs. A total of 24 clean male Sprague Dawley rats were randomly divided into 3 groups： A group （n=12）, transplanted NSC suspension in rat brain after facial nerve injury; B group （n=6）, transplanted nutrient solution of NSCs in rat brain after facial nerve injury; C group （n=6）, transplanted NSCs suspension in normal rat brain. METHODS： Rat models of facial nerve division were established in the groups A and B. 1 week following model induction, NSC stereotaxis transplantation was performed, and the injection point was 11.30 mm to pone of the anterior fontanelle, 9.00 mm to dorsal side, at the median line. 10 μL transgenic NSC suspension （5×10^6 cells） was infused into rats of the groups A and C. 10 μL NSC medium was infused into rats of the group B. Frozen sections of brain tissues were prepared 4 weeks later. MAIN OUTCOME MEASURES： Fluorescence microscope of the histological sections was employed to observe the survival of GFP-positive cells at grafting region and migration of GFP-positive cells in the area around facial nucleus of injured side facial nerve. RESULTS： At grafting region： GFP-positive cells were observed both in groups A and C, and some of these cells were in blood vessel. No GFP-positive cell was observed in group B. tn the area around facial nucleus： In group A, GFP-positive cells were observed migration to the area around facial nucleus of injured side facial nerve; No GFP-positive cell was observed migration to the area around facial nucleus of healthy side facial nerve. In groups B and C, no GFP-positive cell was observed migration to the area around both facial nuclei. CONCLUSION： NSCs could migrate into the area around facial nucleus of injured side facial nerve after transplanted into rat brains.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第19期3771-3775,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
