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

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 (HPL C)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 HPL C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 6 0 k D的蛋白质主带。　结论　结合生物活性检测结果 ,分子量约 6 0 k D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

THE ISOLATION AND BIOACTIVE ASSAY OF NEUROTROPHIC ACTIVE SUBSTANCE DERIVED FROM SPINAL DORSAL HORN OF SPARED ROOT RAT

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.