肝源性肝细胞再生刺激因子的纯化及理化、生物学特性的研究

THE PURIFICATION OF HEPATIC STIMULATOR SUBSTANCE AND ITS PHYSICOCHEMICAL AND BILOLGIACAL CHARACTERISTICS

为更深入了解肝细胞再生刺激因子（ＨＳＳ）的理化及生物学特性，探讨其本质，作者取杂种雄性乳猪，经预先肝切除后取残肝，分别经匀浆、９５℃加热、高速离心、酒精沉淀、凝胶过滤、Ａｍｉｃｏｎ超滤、ＦＰＬＣ层析等一系列纯化过程，并分别进行蛋白定量及生物活性检测。结果显示ＨＳＳ纯化后，蛋白含量降为１／８０．特异性活性提高了９５０倍左右。分子量同定为１３，２００Ｄａ，ＨＳＳ对胰蛋白酶敏感，表明ＨＳＳ是一种具器官特异性，而无种族特异性的热稳定性小分子蛋白。

In order to further elaborate the physiochemical and biological characteristicsof hepatic stimulator substance ( HSS)for better underst anding of the mechnisms ofliver regeneration,a male hybrid piglet was hepatectomied. Forty-eight hours later,the liver remnant was excised and homogenized. The subsequent purification steps in- clude heat,precipitating with ethanol ,high speed centrifuging,passing through Se-phadex G100 colum, ultrafiltrating through the Amicon membrane,and finally byFPLC. The active fractions from all purification steps were determined fortheir pro- tein content as well as their biological activity. The results showed a 500- fold in- crease in purity of HSS with a 950-fold increase in specific activity. SDS-PAGE reveal- ed one major band at 13200Da. HSS is heat resistant, trypsin sensitive,organ spe- cific but not species specific.