|
备用根大鼠脊髓后角组织神经营养活性物质的分离纯化和生物活性测定 |
| |
张媛媛 曾园山 顾熊飞 陈穗君
【摘要】 目的 进一步分离纯化备用根大鼠部分去传入纤维支配(手术侧)的脊髓后角组织提取液，以获得某种神经营养活性物质。 方法 用Superdex G-75 prep grade凝胶层析、高效液相色谱(HPLC)层析和细胞培养等技术分离和检测神经营养活性物质。 结果 手术侧脊髓后角组织提取液经Superdex G-75 prep grade凝胶层析和HPLC层析后，得到的A峰洗脱液具有促进体外培养鸡胚背根节(DRG)分离神经元存活及其神经突起生长的神经营养活性作用。经SDS-聚丙烯酰胺凝胶电泳(PAGE)显示，A峰洗脱液呈现一条分子量约为60 kD的蛋白质主带。 结论 结合生物活性检测结果，分子量约60 kD的蛋白质可能是手术侧脊髓后角组织的神经营养活性物质。
【关键词】 备用根模型；细胞培养；凝胶层析；高效液相色谱；神经营养活性物质
【中图分类号】 Q189 【文献标识码】 A 【文章编号】 0529-1356(2000)04-322
THE ISOLATION AND BIOACTIVE ASSAY OF NEUROTROPHIC
ACTIVE SUBSTANCE DERIVED FROM SPINAL DORSAL HORN
OF SPARED ROOT RAT
ZHANG Yuan-yuan ZENG Yuan-shan CHEN Sui-jun
（Division of Neurosciences, Department of Histology and Embryology）
GU Xiong-fei
（Department of Biochemistry, Sun Yat-Sen University of Medical Sciences, Guangzhou 510089,China）
【Abstract】 Objective The extract of spinal dorsal horn of partial deafferentation(operated side) in spared root rat was isolated and purified further to look for some neurotrophic active substance. Methods Neurotrophic active substance was isolated and analysed by Superdex G-75 prep grade gel chromatography, high performance liquid chromatography (HPLC) and cell culture, ect. Results Peak A eluate was obtained from the extract of spinal dorsal horn of operated side through Superdex G-75 prep grade gel chromatography and HPLC. The peak A eluate played a neurotrophic active role in promoting survival and neurite growth of dissociated neurons of chick embryonal dorsal root ganglion(DRG) in vitro. According to the analysis of SDS-polyacrylamide gel electrophoresis (SDS-PAGE), peak A eluate presented a main protein band with a molecular weight of 60kD. Conclusion The 60kD protein may be a neurotrophic active substance of spinal dorsal horn of operated side by the result of bioactive assay.
【Key words】 Spared root preparation; Cell culture; Gel chromatography; High performance liquid [1] [2] 下一页
上一个医学论文： 晚期糖化终产物对糖尿病大鼠主动脉平滑肌细胞的影响
下一个医学论文： 微囊化转TH基因人成纤维细胞脑内移植治疗帕金森病大鼠模型的研究
|
|
|
|
|
|
|
您现在的位置： 绿色健康网 >> 医学论文 >> 基础医学论文 >> 正文