小牛HSS中有效成分分离及其生物活性的初步研究

PARTIAL PURIFICATION OF THE cHSS EFFECTIVE COMPOSITIONS AND THEIR EFFECTS ON THE NORMAL LIVER AND HEPATOMA CELLS

新生小牛肝细胞刺激物质（ｃＨＳＳ）经超滤、乙醇沉淀、离子交换层析、分子筛层析等步骤分离，获得肝细胞生长正调控因子（ｃＨＳＳα），负调控因子（ｃＨＳＳβ），并观察了它们对肝癌细胞、原代肝细胞的影响。结果表明：ｃＨＳＳα能促进这两种肝细胞的ＤＮＡ合成，其半数有效剂量ＥＤ５０值为３０ｎｇ／ｍｌ，ＳＤＳ－ＰＡＧＥ凝胶电泳显示其分子量为１３０００；而ｃＨＳＳβ明显抑制肝癌细胞的ＤＮＡ合成，其半数抑制剂量ＩＤ５０值为１３０ｎｇ／ｍｌ。

Hepatic stimulate substance is a unique peptide found in the liver of weanling and regenerating,but not normal,adults mammal.It induces liver growth both in vivo and in vitro .It is organ specific but species nonspecific.It also inhibits the hepatoma cell growth in high dose.The hepatocyte growth factor,cHSSα,and the hepatoma cell inhibitor,cHSSβ have been partial purified from cHSS by successive steps,involving ultrafitration,ethanol precipitation,DEAE and Sephadex chromatography.Their bioactivity have been also investigated using the method of 3H TdR incorporation.The results show that the cHSSα,a protein with molecular weight about 13 000,can stimulate both the DNA synthesis of the rat normal liver cell and the human hepatoma cell,and its ED 50 is 30ng/ml.The results also show that the cHSSβ,a protein with molecular weight less than 3 000,can inhibit the DNA synthesis of human hepatoma cell and its ID 50 is 130ng/ml,but it has no inhibiting effect on normal rat liver cell.