Gohda E, Tsubouchi H, Nakayama H, Hirono S, Sakiyama O, Takahashi K, Miyazaki H, Hashimoto S, Daikuhara Y
Department of Biochemistry, Kagoshima University Dental School, Japan.
J Clin Invest. 1988 Feb;81(2):414-9. doi: 10.1172/JCI113334.
Human hepatocyte growth factor (hHGF) has been purified approximately 209,000-fold with 18% yield from plasma of a patient with fulminant hepatic failure. The purification involves heat treatment of plasma, ammonium sulfate precipitation, and chromatography on Affi-Gel Blue, heparin-Sepharose, and hydroxylapatite. Purified hHGF shows several bands with molecular weights between 76,000 and 92,000. Each band shows growth-stimulating activity on cultured 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 hepatocytes is half-maximal at 3.5 ng/ml. hHGF stimulates proliferation of cultured hepatocytes more effectively than human epidermal growth factor (hEGF) or insulin, and the effect of hHGF is additive or synergistic with the maximal effects of hEGF and insulin. These results suggest that hHGF is a new growth factor which is different from hEGF.
人肝细胞生长因子（hHGF）已从一名暴发性肝衰竭患者的血浆中纯化出来，纯化倍数约为209,000倍，产率为18%。纯化过程包括血浆的热处理、硫酸铵沉淀以及在Affi-Gel Blue、肝素-琼脂糖和羟基磷灰石上进行层析。纯化的hHGF显示出几条分子量在76,000至92,000之间的条带。每条条带对培养的肝细胞均显示出生长刺激活性，且该活性与条带强度成正比。用2-巯基乙醇还原样品后，SDS-PAGE产生两条分子量分别为31,500 - 34,500和54,000 - 65,000的链。hHGF对肝细胞DNA合成的作用在3.5 ng/ml时达到最大效应的一半。hHGF比人表皮生长因子（hEGF）或胰岛素更有效地刺激培养的肝细胞增殖，并且hHGF的作用与hEGF和胰岛素的最大效应具有相加或协同作用。这些结果表明hHGF是一种不同于hEGF的新生长因子。
