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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(1μL/min)。对照组注射生理盐水10μL。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,P<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,P<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。
BACKGROUND: Olfactory ensheathing cells, as a kind of regenerative and self-derived transplanted cells, are widely concerned in the treatment of spinal cord diseases. The transplantation of intracerebral hemorrhage is yet to be further accumulated. OBJECTIVE: To observe the proliferation of neural precursor cells in rats with cerebral hemorrhage after transplantation of olfactory ensheathing cells and evaluate the effect of olfactory ensheathing cells transplantation on intracerebral hemorrhage in rats. Design: Complete randomized controlled experiment. SETTING: Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was performed at the Center of Clinical Neurology, Tongji Medical College, Huazhong University of Science and Technology from March 2002 to March 2003. Thirty - two healthy male Wistar rats were randomly divided into two groups of 16. Olfactory ensheathing cells transplantation group in the production of rat caudate nucleus hemorrhage model day 3, take 10 μL of olfactory ensheathing cell suspension to rat brain uniform injection (1μL / min). The control group was injected with normal saline 10μL. Methods: The neurological scores of rats were respectively taken before and 3, 7, 14 and 30 days after transplantation. On the first day after the model was made, one rat was taken from each of two groups to prepare the brain tissue sections. The axons of the axons were observed with myelin solid blue staining and the nerve fibers were observed with the neurofibrillary silver staining. On the 3rd, 7th, 14th and 30th day after transplantation, one rat was taken from each group and paraffin sections were made. The survival, migration and proliferation of neural precursor cells were observed after transplantation of olfactory ensheathing cells and the neural precursor cells were counted. MAIN OUTCOME MEASURES: ①Nerve cell axon myelin and nerve fibers were observed after intracerebral hemorrhage in both groups. ② The number of neural precursor cells on the 3rd, 7th, 14th and 30th day after intracerebral hemorrhage in both groups were counted. ③ The neurological deficit score of the 3, 7, 14 and 30 days after intracerebral hemorrhage in both groups. Results: All 32 rats were involved in the experiment. ①On the 30th day after intracerebral hemorrhage, the numbers of OECs in perihematoma and hematoma in the hematoma were significantly higher than those in the control group. ②Nerve precursor cell count after intracerebral hemorrhage in both groups: On the 7th, 14th and 30th day after ICH, the number of neural precursor cells in the olfactory ensheathing cell transplantation group was significantly more than that in the control group (41.1 ± 2.4) / field , (34.5 ± 1.2) / field of vision, (43.6 ± 1.2) / field of vision, (37.2 ± 2.0) / field of vision, (19.3 ± 1.0) / field of vision, (14.2 ± 0.4) ~ 4.02, P <0.05)]. (3) The score of neurological deficit after intracerebral hemorrhage in both groups was significantly lower than that of the third day [(2.21 ± 0.20), (1.50 ± 0.21), (2.74 (0.96 ± 0.12) and (2.76 ± 0.20) points on the 30th day after cerebral hemorrhage, respectively (t = 2.06,3.27, P <0.05) = 2.47, P <0.05)]. Conclusion: Olfactory ensheathing cells can increase the number of endogenous neural precursor cells. Olfactory ensheathing cells can promote axonal regeneration of nerve cells in the brain, make it remyelination and establish synaptic connection, restore its motor function, and then accelerate the repair of injured tissue.