[Purification and partial characterization of hepatocyte growth factor from plasma of patients with fulminant hepatic failure].
[Purification and partial characterization of hepatocyte growth factor from plasma of patients with fulminant hepatic failure].
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暴发性肝衰竭患者血浆中肝细胞生长因子的纯化及部分表征
DOI：
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
O. Sakiyama
中科院分区：
文献类型：
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作者：
O. Sakiyama
人肝细胞生长因子（hHGF）已从暴发性肝功能衰竭患者的血浆中纯化。纯化程序包括五个步骤，血浆热处理、硫酸铵沉淀以及Affi-Gel Blue、肝素-Sepharose和羟基磷灰石层析。从患者的930 ml血浆开始，已获得37微克hHGF，代表209，000倍纯化和18%产率。纯化的hHGF显示分子量在76，000和92，000之间的几条带。两个主要条带的分子量为79，000和86，000。每个条带显示对培养的大鼠肝细胞的生长刺激活性，其与条带的强度成比例。用2-巯基乙醇还原样品后，SDS-PAGE产生分子量为31，500 - 34，500和54，000 - 65，000的两条链。hHGF对肝细胞DNA合成的影响在3.5 ng/ml时达到半数最大。胰岛素样生长因子-I和-II，成纤维细胞生长因子，血小板衍生生长因子和转化生长因子-β，这是已知的生长因子，在培养的大鼠肝细胞的DNA合成没有刺激作用。hHGF比人表皮生长因子（hEGF）或胰岛素更有效地刺激培养的大鼠肝细胞增殖，并且hHGF的作用与hEGF和胰岛素的最大作用是额外的或协同的。用这三种生长因子（胰岛素、hHGF和hEGF）的培养物的标记指数为85%，表明大多数肝细胞被诱导增殖。提示hHGF是一种不同于hEGF的新的生长因子。
Human hepatocyte growth factor (hHGF) has been purified from plasma of a patient with fulminant hepatic failure. The purification procedure consists of five steps, heat treatment of plasma, ammonium sulfate precipitation, and chromatography on Affi-Gel Blue, heparin-Sepharose, and hydroxylapatite. Starting with 930 ml of plasma from a patient, 37 micrograms of hHGF has been obtained, representing a 209,000-fold purification and a 18% yield. Purified hHGF shows several bands with molecular weights between 76,000 and 92,000. The two major bands had molecular weights of 79,000 and 86,000. Each band shows growth-stimulating activity on cultured rat hepatocytes which is proportional to the intensity of the band. After reduction of the sample with 2-mercaptoethanol, SDS-PAGE yields two chains with molecular weights of 31,500-34,500 and 54,000-65,000. The effect of hHGF on DNA synthesis by hepatocyte is half-maximal at 3.5 ng/ml. Insulin-like growth factor -I and -II, fibroblast growth factor, platelet-derived growth factor and transforming growth factor-beta, which are known as growth factors, had no stimulatory effect on DNA synthesis in cultured rat hepatocytes. hHGF stimulates proliferation of cultured rat hepatocytes more effectively than human epidermal growth factor (hEGF) or insulin, and the effect of hHGF is additional or synergistic with the maximal effects of hEGF and insulin. Labeling index of cultures with these three growth factor (insulin, hHGF and hEGF) was 85%, showing that most hepatocytes were induced to proliferate. These results suggest that hHGF is a new growth factor which is different from hEGF.