嗅鞘细胞移植后脑出血大鼠神经前体细胞增殖及神经功能评分变化

背景嗅鞘细胞作为一种可再生且自体取材的移植细胞,在脊髓疾病移植治疗中受到广泛关注,关于脑出血的移植治疗目前有待实验结果的进一步积累.目的观察嗅鞘细胞移植后脑出血大鼠神经前体细胞的增殖,评估嗅鞘细胞移植治疗大鼠脑出血的疗效.设计完全随机对照实验.单位华中科技大学同济医学院附属同济医院神经内科.材料实验于2002-03/2003-03在华中科技大学同济医学院临床神经研究中心完成.选择健康雄性Wistar大鼠32只随机分为2组,每组16只.嗅鞘细胞移植组在制作大鼠尾状核出血模型第3天时,取10μL嗅鞘细胞悬液向大鼠脑内匀速注射(1 μL/min).对照组注射生理盐水10 μL.方法大鼠在移植前,移植后第3,7,14,30天分别进行神经功能评分.模型制作后第1天在两组中各取1只大鼠,制备脑组织切片,神经元轴突髓鞘观察采用髓鞘固蓝染色,神经纤维观察采用神经纤维嗜银染色.各组移植后第3,7,14,30天各取1只大鼠,制作石蜡切片,观察嗅鞘细胞移植后存活、迁徙及神经前体细胞的增殖情况,并进行神经前体细胞计数.主要观察指标①两组大鼠脑出血后神经元轴突髓鞘和神经纤维观察.②两组大鼠脑出血后第3,7,14,30天神经前体细胞计数.③两组大鼠脑出血后第3,7,14,30天时神经功能缺损评分.结果32只大鼠均进入实验分析.①脑出血后第30天时血肿周边及血肿灶中嗅鞘细胞计数移植组的髓鞘化数量和神经纤维数量明显多于对照组.②两组大鼠脑出血后神经前体细胞计数脑出血后第7,14,30天,嗅鞘细胞移植组的神经前体细胞数明显多于对照组(41.1±2.4)个/视野,(34.5±1.2)个/视野;(43.6±1.2)个/视野,(37.2±2.0)个/视野;(19.3±1.0)个/视野,(14.2±0.4)个/视野,(t=2.42～4.02,＜0.05)].③两组大鼠脑出血后神经功能缺损评分嗅鞘细胞移植组在脑出血后第14,30天时明显低于第3天(2.21±0.20)分,(1.50±0.21)分,(2.74±0.21)分,(t=2.06,3.27,＜0.05)],对照组只在脑出血后第30天时低于第3天(1.96±0.12)分,(2.76±0.20)分,(t=2.47,P＜0.05)].结论①嗅鞘细胞有增加内源性神经前体细胞数量的作用.②嗅鞘细胞可促进脑内神经细胞轴突再生,使其重新髓鞘化并建立突触联系,恢复其运动功能,进而加快损伤组织的修复.

Proliferation of neural progenitor cells and evaluation of neurologic function in cerebral hemorrhagic rats after transplantation of olfactory ensheathing cells

BACKGROUND: Being a kind of regenerative and auto-transplanting cell, olfactory ensheathing cell (OEC) has been extensively concerned on transplantation treatment for spinal disease. Concerning to the transplantation in treatment of cerebral hemorrhage, it is expected a further accumulation of experimental results at present.OBJECTIVE: To observe the proliferation of neural progenitor cells in cerebral hemorrhagic rats after OEC transplantation and to evaluate the therapeutic effects of OEC transplantation on cerebral hemorrhage.DESIGN: Completely randomized controlled experiment.SETTING: Department of Neurology of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Research Center for Clinical Neurology , Tongji Medical College of Huazhong University of Science and Technology from March 2002 to March 2003. Thirty-two healthy male Wistar rats were employed and randomized into 2 groups, 16 rats in each. In OEC transplantation group, on the 3rd day of modeling hemorrhage of caudate nucleus, OEC suspension 10 μL was injected evenly in the brain of rat (1 μL/min). In the control group, physiological saline 10 μL was injected.METHODES: Neural function evaluation was done before transplantation,on the 3rd, 7th, 14th and 30th days after transplantation successively. On the first day after modeling, 1 rat was collected from each of two groups to prepare brain tissue section. Myelin sheath blue staining was used for observation of neuronal axonal myelin sheath. Never fiber argentophil staining was used for observation of never fiber. One rat was collected from each of two groups on the 3rd, 7th, 14th and 30th days after transplantation successively to prepare paraffin section. The survival and migration after OEC transplantation as well as proliferation of neural progenitor cell were observed.The count of neural progenitor cell was recorded.myelin sheath and nerve fiber after cerebral hemorrhage in rats of two function deficits on the 3rd, 7th, 14th and 30th days after cerebral hemorrhage in rats of two groups.around and in hematoma on the 30th day after cerebral hemorrhage: In transplantation group, myelinated amount and nerve fiber amount were cell after cerebral hemorrhage in rats of two groups: on the 7th, 14th and 30th days after cerebral hemorrhage, the amount of neural progenitor cell in OEC transplantation group was more remarkably than that in the control group (41.1 ±2.4)pcs/vision field, (34.5 ±1.2)pcs/vision field; (43.6±1.2)pcs/vision rield, (37.2±2.0)pcs/vision field; (19.3±1.0)pcs/vision rield, ( 14.2±0.4)pcs/videficits after cerebral hemorrhage in rats of two groups: In OEC transplantation group, on the 14th and 30th days, the evaluation was lower remarkably than the 3rd day (2.21 ±0.20)scores, (1.50±0.21)scores, (2.74±0.21)scores, (t=2.06, 3.27, P ＜ 0.05)]. In the control group, that on the 30th day after cerebral hemorrhage was lower than that on the 3rd day (1.96±0.12)scores ,(2.76±0.20) scores, (t=2.47, P ＜ 0.05 )].tion of intracerebral nerve cell, re-myelination and building-up synaptic system so as to recover the motor function and accelerate repair of injured tissue.