胚胎肢芽内神经营养活性物质的提纯及其生物活性的研究

Extracting and purification neurotropins from embryonic limb buds and study of their activity

目的研究从胚胎肢芽内提取和纯化具有神经营养活性的物质。方法剖腹取胚龄为１０～１５天的ＳＤ大鼠胚胎，将其肢芽连同基部剪下、剪碎。匀浆离心后取上清液超滤，分别收集大于１００００ＫＤ的组分和小于１００００ＫＤ的组分。然后将大于１００００ＫＤ的组分用高速蛋白液相层析系统分离，得５个蛋白组分，即＞３０００００ＫＤ、２１００００ＫＤ、９５０００ＫＤ、３４０００ＫＤ和２２０００ＫＤ组分。再将这５个组分连同小于１００００ＫＤ的组分，分别加至胚鼠背根神经节感觉神经元的培养基中，培养２４小时后用ＭＴＴ法测定神经元的活性。结果９５０００ＫＤ、３４０００ＫＤ和２２０００ＫＤ３个组分对神经元有营养活性。其中２２０００ＫＤ的活性较弱，９５０００ＫＤ和３４０００ＫＤ的活性较强。此外，２２０００ＫＤ、３４０００ＫＤ和９５０００ＫＤ合在一起的活性明显强于任何单一组分。结论胚胎肢芽内含有较强的促进神经元活性的神经营养活性物质，为神经营养因子提供了新的来源。

Objective To extract and purify neurotropins from limb embryonic buds and study their bioactivity. Methods SD rats embryoes at the age of 10～15 days were obtained by cesarean section.Limb buds with their bases were cut off,and put in homogenizer for homogenating.After centrifugation, the supernate was ultrafiltrated and passed HPLC molcular column. The proteins in limb buds were then separately components of < 10 000 Results The components of 22 000 Conclusions Neurotrophic substances with high activity are present in limb buds,and limb buds may become a new source of neurotrophic factors (NTFs).