生理学报Acta Physiologica Sinica, June 25, 2003, 55(3)
研究论文
胚胎神经干细胞移植及胶质细胞源性神经营养因子对大鼠脊髓损伤的修复作用
孙勇1, 侍坚1, 富赛里1, 陆佩华1，*, 徐晓明1,2
1上海第二医科大学神经生物学实验室, 上海 200025;
2美国肯塔基脊髓损伤研究中心, 美国路易斯维尔大学医学院神经外科, 路易维尔市, 肯塔基州 40292
摘要: 将胚胎神经干细胞(neural stem cells, NSCs)移植至成年大鼠损伤的脊髓, 观察移植后NSCs的存活、 迁移以及损伤后的功能恢复。实验结果显示: 动物NSCs移植4周后, 斜板实验平均角度和运动评分结果比对照组均有明显增高(P<0.05), 而脊髓损伤(spinal cord injury, SCI)处的空洞面积显著减小(P<0.05); 在NSCs中加入胶质细胞源性的神经营养因子(glial cell line-derived neurotrophic factor, GDNF)后, 上述改变更加显著。移植后的NSCs不仅能存活, 而且向损伤的头端和尾端迁移达3 mm之远。这些结果表明, 移植的NSCs不仅可以存活、 迁移, 还可减小 SCI空洞面积, 促进动物神经功能的恢复; 此外, 我们的结果还表明GDNF对SCI功能恢复有促进作用。
关键词: 脊髓损伤; 胚胎神经干细胞; 移植; 胶质细胞源性的神经营养因子
中图分类号: Q426; R743.31
Effects of
embryonic neural stem cells and GDNF in the repair of spinal cord injury
SUN Yong1, Shi Jian1, Fu Sai-Li1, Lu Pei-Hua1, Xu Xiao-Ming1,2
1Department of Neurobiology , Shanghai Second Medical University, Shanghai 200025;
2Kentucky Spinal Cord Injury Research Center, Department of Neurological Surgery, University of Louisville, 511S. Floyd Street MDR616, Louisville, KY40292
Abstract: The ability of implanted embryonic neural stem cells (NSCs) to improve survival, migration, and functional recovery following a compression spinal cord injury (SCI) was tested in adult rats. NSCs were isolated from E14-16 rat cerebral cortex and SCI was produced by using an aneurysm clip applicator applied to the 8th thoracic spinal cord according to method of Dolan and Tator. Two weeks after the injury, NSCs (4 μl of 1×104 cells/μl) were injected into the lesion site., The grafted NSCs were noted to survive and integrate with the host spinal cord 1 month after transplantation, which was demonstrated by the presence of Hoechst 33342 (a nuclear dye) pre-labeled NSCs within and surrounding the lesion site. Some of these cells remained undifferentiated and were stained with nestin, a marker for NSCs. Transplanted NSCs migrated for at least 3 mm from the injury epicenter towards both the rostral and caudal directions. Significant reduction in the lesion area (P<0.05) and improvement in inclined plane (P<0.05) and BBB locomotor rating scale (P<0.05) were found in the cases that received implantation of NSCs, as compared with those that received vehicle injection. More importantly, when glial cell line-derived neurotrophic factor (GDNF; 1.5 μg/μl) was added to the transplants, further reduction in lesion area (P<0.01) and improvement in the function were observed in the combined treatment group as compared with the vehicle infused group. Our results suggest that intraspinal treatment with NSCs and GDNF synergistically reduced lesion size and improved functional outcome after a compressive SCI in adult rats.
Key words: spinal cord injury; neural stem cell; transplantation; glial cell line-derived neurotrophic factor