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我室新近的研究发现,部分去传入纤维支配的脊髓后角组织提取液含有神经营养活性组份,其分子量约在40KD～78KD之间,但不清楚是那种分子量的蛋白质具有神经营养活性作用.为了寻找这种物质,本研究应用Superdex G-75prep grad凝胶层析、高效液相色谱(HPLC)层析和细胞培养等技术和方法,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,以期获得某种神经营养活性物质.用SD雄性大鼠100只,切除其一侧L_1-L_3、L_5和L_6背根,仅保留L_4背根(此动物为备用根模型大鼠),术后5天处死动物.分别切取L1_～L_6节段的手术侧和非手术侧脊髓后角组织,制备成提取液、凝胶层析洗脱液和PHLC层拆洗脱液.结果发现,手术侧脊髓后角组织提取液具有神经营养活性作用,将该提取液经Superdex G-75prep grade凝胶层析和PHLC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-聚丙烯酸胺凝胶电泳(PAGE)显示,A峰洗脱液呈现一条分子量约为60kD的蛋白质主带.结合生物活性检测结果,我们推测分子
Recent studies in our laboratory found that some of the extracts from the dorsal horn of the spinal cord dominating the afferent fibers contain neurotrophic components with a molecular weight of about 40 kD to 78 kD but it is unclear which protein of that molecular weight has neurotrophic activity In order to find out the substance, this study used Superdex G-75prep grad gel chromatography, high performance liquid chromatography (HPLC) chromatography and cell culture techniques and methods to further purify the spare root rats to some of the imported fibers Controlled spinal cord posterior horn tissue extract (surgery side), in order to obtain some neurotrophic substances.Using 100 SD male rats, the L_1-L_3, L_5 and L_6 dorsal roots were excised on one side and only the L_4 dorsal root The animals as spare root model rats) and animals were sacrificed 5 days after operation.The surgical and non-surgical spinal dorsal horn tissues of L1_ ~ L_6 segments were cut and extracted into extracts, gel chromatography eluate and PHLC Layer disassembled eluate.It was found that the surgical side of the spinal cord posterior horn tissue extract has neurotrophic activity role in the extract by Superdex G-75prep grade gel chromatography and PHLC chromatography, the resulting A peak eluate Promote chick embryo back culture in vitro (DRG) to separate the neuronal survival and the neurotrophic activity of its neurite growth.A SDS-polyacrylamide gel electrophoresis (PAGE) showed that the A peak eluted protein showed a protein main band with a molecular weight of about 60 kD. Bioactivity test results, we speculate molecules