雪旺细胞源神经营养蛋白的纯化及其体外生物活性的研究

THE PURIFICATION OF SCHWANN CELLS-DERIVED NEUROTROPHIC SUBSTANCES AND STUDYING ON IT'S NERUOBIOLOGICAL ACTIVITY

目的：纯化和研究源于雪旺细胞质的运动神经营养因子。方法：２００条预损伤２周后的ＳＤ大白鼠坐骨神经，用酶消化结合差速粘附法纯化并收集其中的雪旺细胞；超声粉碎雪旺氏细胞，离心，收集上清液再经起滤，过高效液相分子柱，雪旺细胞上清液蛋白分为分子量分别为小于１０ＫＤ，２６ＫＤ，５０ＫＤ，７５ＫＤ，及大于２００ＫＤ五个组分，分别和胎鼠脊髓运动神经元联合培养２４小时，用ＭＴＴ法测定神经元活性。结果：２６ＫＤ，５０ＫＤ，和７５ＫＤ蛋白组分对脊髓运动神经元有维持成活的营养作用，结论：雪旺细胞可合成和分泌三种运动神经营养蛋白，分子量分别为２６ＫＤ，５０ＫＤ和７５ＫＤ。

Objective:To purify and study Schwann cells derived motoneurotrophic factors(MNTFS).Methods:Schwann-cells (SCs)were purified and collected by the methods of enzyme di- gesting and speed difference adhesion from two hundreds sciatic nerves of adult SD rats after two weeks,These SCs were ultrasonicated and centrifuged and the supernate were collected.After ultrafil- trated and passing through HPLC molecular columm,the proteins in supernate were separated into five components. The molecular weight is seperatedly<10KD,26KD,50KD,75KD,and>200KD。They were separatedly added into the medium of rat embryonic spinal cord motoneurons,and after culturing 24 hours,using MTT method examined the living activity of the neurons,Results:The components 26KD,50KD and 75KD have the trophic effects of maintaining survival to spinal cord motoneu- rons,Conclusion:SCs could synthesize and secrete three kinds of MNTFs and the molecular weight seperatedly are 26KD,50KD and 75KD.