The Study of the Hepatotrophic Factor in Seva from Patients with Fulminant Gepatic Failure. -its purification and clinical significarce-

按照最初的计划，我们对人肝卵细胞生长因子（hHGF）进行了研究，得到了以下结果：(1)用原代培养的成年大鼠肝细胞测定肝细胞的生长情况。20例暴发性肝衰竭患者血清或血浆的碘含量。“急性和慢性”患者的血清对DNA合成的刺激活性明显高于正常人血清的平均值约5-50倍。(2)从暴发性肝衰竭患者的加热血浆中纯化出约21万倍的hHGF。提纯涉及等离子体硫酸铵沉淀的热处理。和Affi-Gel Blue的柱层析。hepalin-Sopharose。和羟基磷灰石。(3)纯化后的hHGF在SDS-PAGE上显示出分子量约为80,000的条带。每个波段对培养的肝细胞显示出刺激生长的活性，这与波段的强度成正比。用2-巯基乙醇还原样品后。SDS-PAGE得到两条分子量约为30000和60000的链。(4) hHGF比人表皮生长因子（hHGF）和胰岛素更能促进培养大鼠肝细胞的生长。hHGF的作用与胰岛素的最大作用具有叠加和协同作用。分别。这些结果表明，hHGF是一种不同于hHGF的新型生长因子。(5)。以纯化的hHGF为抗原制备hHGF单克隆抗体。目前我们正在研究利用该抗体产生hHGF的器官。

According to the initial plan, we studied the human hepatooytc growth factor (hHGF) and obtained the following results.(1) Liver cell growth was measured using adult rat liver celis in primary culture. inetoon of twenty sera or plasma from patients with fulminant hepatic failure. and serum from a patient with "acute on chronic" showed marked stimulatoy activity on DNA synthesis at about 5-50 times higher than an avarage value of normal human sera.(2) hHGF was purified at about 210,000-fold from heated plasma from patients with fulminant hepatic failure. The pulification invelves heat treatment of plasma ammonium sulfate precipitation. and column chromatographies on Affi-Gel Blue. hepalin-Sopharose. and hydroxylapatite.(3) Pulified hHGF shows several bands with molecular weights of about 80,000 on SDS-PAGE. Each bands shows growth-stimulating activity on cuitured hepatocytes which is propotional to the intensity of the band. After reduction of the sample with 2-mercptoethanol. SDS-PAGE yield two chaines with molecular weights of about 30,000 and 60,000.(4) hHGF stimulates growth of cultured rat hepatocytes more effectively than human epidermal growth factor(hHGF) or insulin. and the effect of hHGF is additive and synergistic with the maximal effects of hHGF and insulin. respectively. These results suggest that hHGF is a new growthfactor which is different from hHGF.(5) Recently. a monoclonal antibody to hHGF was raised using purified hHGF as antigen. At present we are studying the organ producing hHGF using with this antibody.