论文部分内容阅读
目的 分离纯化吗啡备用根大鼠脊髓组织提取液 ,以期获得某些神经营养活性物质。 方法 应用SephacrylS 2 0 0HR凝胶层析、高效液相色谱 (HPLC)和组织培养等技术分离和检测神经营养活性物质。 结果 备用根大鼠脊髓组织提取液能够促进体外培养的鸡胚背根节 (DRG)神经突起的生长 ;吗啡作用的大鼠脊髓组织提取液也具有同样的作用 ,但备用根大鼠和吗啡作用的大鼠脊髓组织提取液在促神经突起生长作用方面并没有明显的差异。吗啡备用根大鼠脊髓组织提取液具有明显的神经营养活性作用。吗啡备用根大鼠脊髓组织提取液的SephacrylS 2 0 0HR凝胶层析Ⅱ峰洗脱液和Ⅳ峰洗脱液能够促进DRG神经突起的生长。经SDS PAGE分析 ,Ⅱ峰洗脱液呈现 1条分子量约为 6 5kD的蛋白质主带 ,Ⅳ峰洗脱液的蛋白质成分较为复杂。应用HPLC对凝胶层析Ⅳ峰洗脱液作进一步的分离 ,发现HPLCA峰洗脱液能够促进DRG神经突起的生长。经SDS PAGE显示 ,A峰洗脱液的两条蛋白质主带分子量分别为 30kD和 18kD。 结论 吗啡备用根大鼠脊髓组织提取液中具有神经营养活性作用的物质可能是分子量约为 6 5kD、30kD和 18kD蛋白质
OBJECTIVE To isolate and purify spinal cord tissue extracts of morphine spare root in order to obtain certain neurotrophic substances. Methods Separation and detection of neurotrophic substances by Sephacryl S 200HR gel chromatography, high performance liquid chromatography (HPLC) and tissue culture techniques. Results Spinal cord tissue extracts of spare root rats could promote the growth of chick embryo dorsal root ganglion (DRG) neurons in vitro. Morphine-treated rat spinal cord tissue extract also had the same effect, but the effects of morphine Of rats spinal cord tissue extract in the promotion of neurite outgrowth role and no significant differences. Spinal cord extract of morphine spare root has obvious neurotrophic activity. Morphine Spinal Root Separation from Spinal Cord Tissue Extract Sephacrylsol 200HR Gel Elution Ⅱ and Ⅳ Peak Eluent can promote the growth of DRG neurite. According to SDS PAGE analysis, the peak of Ⅱ peak showed one main protein band with a molecular weight of about 65 kD. The protein fraction of Ⅳ peak eluate was more complicated. The HPLC chromatographic peak eluate was further separated by HPLC, and it was found that the peak HPLCA eluate can promote the growth of DRG neurite. SDS PAGE showed that the main band molecular weights of the two proteins in the peak A eluate were 30 kD and 18 kD, respectively. Conclusion The substance with neurotrophic activity in the spinal cord tissue of morphine spare root may be the protein with the molecular weight of about 65 kD, 30 kD and 18 kD