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An experimental study for the functional recovery of injured spinal motoneurons

脊髓运动神经元损伤功能恢复的实验研究

基本信息

批准号：
09671519
负责人：
FUJIMURA Yoshikazu
金额：
$ 1.73万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671519/
关键词：
spinal cord injury motoneuron nerve growth factor p-75 glial cell line-derived neurotrophic factor choline acetyltransferase コリンアセチル基転移酵素神経栄養因子受容体神経成長因子神経成長因子受容体 Glial cell line-derived neurotrophic factor

项目摘要

We investigated the influences of neurotrophic factor on injured motoneurons in acute spinal cord injury models. Our study consists of the following two experimentsExperiment 1.25 Wister rats were used in this experiment. An incomplete spinal cord injury model was created by placing a 20-g weight on the dura mater at the C6 level for 5 min (n=20) .The cervical spinal cord was remove at post-injury day 3,7,14,28, and frozen sections were stained for p-75 .The number of p-75 positive motoneurons were counted. (Results) There were no p-75 positive motoneurons in the sham group(n=5). In the injury group, p-75 positive motoneurons were recognized at post-injury day 3, highly increased at day 7, and hardly observed by at day 14, 28.Experiment. 2; 20 incomplete spinal cord injury models were created in the same manner. In 10 glial cell line-derived neurotrophic factor(GDNF) was injected into the subarachnoid space immediately after the injury, ,and phosphate buffer saline (PBS) was injected n the other 10 animals. Motor function was evaluated at post-injury day 2 and 7. The spinal cord was removed from both groups 7 days after the injury. The sections were immunofluorescence stained for choline acetyltransferase (ChAT). ChAT fluorescence was measured with a microphotometry system. (Results) Motor function evaluation: Both the GDNF group and the PBS group were able to maintain their trunk on the inclined plane up to a mean angle of 82゜before the injury. On post-injury day 2 the angle at which they, were able to maintain their trunk had decreased to 55゜and 52゜, respectively, and on day 7, to 68゜and 63゜ respectively. The deference on post-injury day 2 was not significant, but a significant deference was observed by day 7. Immunohistochemical findings: The fluorescence values in both injured groups were lower than in the uninjured group, but the reduction in fluorescence that occurred in the GDNF group was a significantly prevented.

本研究探讨了神经营养因子对急性脊髓损伤模型中受损运动神经元的影响。本研究包括以下两个实验：实验一：25只Wister大鼠。在C6水平硬脑膜上放置20 g重物5 min造成不完全性脊髓损伤模型（n=20），分别于伤后3、7、14、28 d取颈髓，冰冻切片进行p-75染色，计数p-75阳性运动神经元数目。（结果）假手术组（n=5）未见p-75阳性运动神经元。损伤组在伤后3天出现p-75阳性运动神经元，伤后7天明显增多，伤后14、28天几乎未观察到。2; 20只不完全性脊髓损伤模型。其中10只动物于伤后即刻蛛网膜下腔注射胶质细胞源性神经营养因子（GDNF），另10只动物于伤后即刻蛛网膜下腔注射磷酸盐缓冲液（PBS）。在损伤后第2天和第7天评价运动功能。损伤后7天，两组均取出脊髓。对切片进行胆碱乙酰转移酶（ChAT）免疫荧光染色。用显微光度系统测量ChAT荧光。（结果）运动功能评价：伤前GDNF组和PBS组均能保持躯干在斜面上的平均角度达82 °。受伤后第2天，他们能够保持躯干的角度分别降至55 °和52 °，第7天分别降至68 °和63 °。损伤后第2天的差异不显著，但到第7天观察到显著差异。免疫组化结果：两个损伤组的荧光值均低于未损伤组，但GDNF组中发生的荧光减少被显著阻止。