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

根据最初的计划，我们对人肝细胞生长因子(HHGF)进行了研究，获得了以下结果：(1)用成年大鼠肝细胞进行原代培养，测定肝细胞的生长。对20例暴发性肝衰竭患者的血清或血浆进行检测。而一名“急慢性”患者的血清对DNA合成的刺激活性约为正常人血清平均值的5-50倍。(2)从暴发性肝功能衰竭患者加热的血浆中纯化的hHGF约为21万倍。制浆涉及血浆硫酸铵沉淀物的热处理。和Affi-Gel Blue的柱层析。肝素-葡萄糖素。(3)打浆后的人肝细胞生长因子在SDS-PAGE上显示多条相对分子质量约为8万的条带。每条带对培养的肝细胞都有刺激生长的活性，与带的强度成正比。样品用2-甲基乙醇还原后。SDS-PAGE显示两条链的相对分子质量分别为30,000和60,000。(4)人肝细胞生长因子(HHGF)比人表皮生长因子(HHGF)或胰岛素更有效地刺激培养的大鼠肝细胞生长。HHGF的作用是相加和协同的，其中hHGF和胰岛素的作用最大。分别进行了分析。这些结果表明，hHGF是一种不同于hHGF的新的生长因子。以纯化的人肝细胞生长因子为抗原，制备了抗人肝细胞生长因子的单抗。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.