备用根大鼠脊髓后角提取液中神经营养活性组分的分离及生物活性测定

ISOLATION AND BIOASSAY OF THE NEUROTROPHIC ACTIVE FRACTION TN TISSUE EXTRACT FROM SPINAL DORSAL HORN OF SPARED ROOT RAT

制作备用根大鼠脊髓后角提取液，取其分子量大于10kD的组分进行SDS-聚丙烯酰胺凝胶电泳分析，发现手术侧和非手术侧的电泳区带数目大致相同，但手术侧样品的第四条蛋白质区带扫描的吸收峰面积百分比大于非手术侧样品的第四条区带，两者在量上可能有差别。将手术侧样品经交联葡聚精G-75凝胶层析，得到两个洗脱峰．第一峰洗脱液有促进体外培养的背根节神经元存活及其突起生长的作用，其电泳分析显示4条主带，分子量在40～80kD之间．实验结果提示部分去传入纤维支配的脊髓后角组织含有神经营养活性物质，该物质的分子量约在40～78kD之间．

Tissue extract frorn spinal dorsal horn of spared root rat was prepared and it's fraction of >l0 kD was analyzed with SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The number of electrophoretic zones of samples was approximately same in operated side and unoperated side, whereas the area percentage of scanning absorption peak of the fourth electrophoretic zone of the sample in operated side was tending towards increasing in comparison with unoperated side. It suggests that the protein content of the fourth zone in operated side varies from that in unoperated side. The sample of operated side was further separated by Sephadex G-75 gel chromatography. Two eluting peaks were obtainted. The survival and neurite-promoting effects on cultured dorsal root ganglion (DRG) neurons of chick embryo were found in the peak l eluate,which showed four electrophoretic zones (molecular weight: 40~78 kD) by SDS-PAGE. The results indicate that partially deafferented spinal dorsal horn contains neurotrophic active substances which may be some proteins of molecular weight 4O^78 kD.