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The Role of a New Hepatocyte Growth Factor in Liver Regeneration

新型肝细胞生长因子在肝脏再生中的作用

基本信息

批准号：
01571206
负责人：
GOHDA Eiichi
金额：
$ 1.34万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01571206/
关键词：
Hepatocyte growth factor Hepatocytes in primary culture Hepatic injury Liver regeneration Carbon tetrachloride

项目摘要

A mouse serum hepatocyte growth-stimulating activity markedly increased 24 h after carbon tetrachloride administration and then decreased. This time course of the activity was paralled with those of serum alanine and aspartate aminotransferses activities. hepatocyte growth-stimulating activity in liver extracts was low in untreated mice, but increased after carbon tetrachloride. The increase was detectable 12 h and marked 24 h after the hepatotoxin. It was maximal 48 h after the administration and then dropped gradually. Liver DNA synthesis began to rise and was highest 36 h and 48 h, respectively, after the hepatotoxin. Thus, the hepatocyte growth-stimulating activity in serum and liver increased prior to liver regeneration in carbon tetrachloride-treated mice. The elevated serum and liver activities may play an important role in promoting proliferation of liver cells in vivo subsequent to hepatic injury. Mouse hepatocyte growth factor (mHGF) was purified about 280,000-fold with 21% yield form the liver supernatant of mice treated with the hepatotoxin by ammonium sulfate fractionation followed by chromatography on Affi-Gel Blue, heparin-Sepharose and S-Sepharose. SDS-PolyacrylAmide Gel Electrophoresis (PAGE) of purified mHGF under nonreducing conditions showed a single band with a Molecular Weight (MW) of 76K, while two bands with MWs of 62K and 31K were detected after SDS-PAGE of reduced mHGF, showing that mHGF is a heterodimer. mHGF stimulated DNA synthesis of cultured hepatocytes as effectively as hHGF. The effect of mHGF was additive with the maximal effects of EGF and acidic FGF and synergistic with that of insulin, but not with the maximal effect of hHGF. Thus, the properties of mHGF are similar to those of hHGF. mHGF may be a hHGF-like factor with some structural differences from hHGF, because the activity of mHGF was not inhibited by rabbit anti-hHGF antiserum which neutralizes the hHGF activity.

四氯化碳给药24 h后，小鼠血清肝细胞促生长活性显著升高，随后下降。该时间过程与血清丙氨酸和天冬氨酸转氨酶活性基本一致。肝提取物刺激肝细胞生长的活性在未处理的小鼠中较低，但在四氯化碳处理后增加。肝毒素作用后12小时可检测到升高，24小时可见。给药后48 h最大，随后逐渐下降。肝毒素作用后36 h和48 h，肝脏DNA合成开始升高，分别达到最高。因此，在四氯化碳处理小鼠的肝脏再生之前，血清和肝脏中的肝细胞生长刺激活性增加。肝损伤后血清和肝脏活性的升高可能在促进肝细胞在体内的增殖中起重要作用。小鼠肝细胞生长因子（mHGF）经硫酸铵分离，再经Affi-Gel Blue、肝素- sepharose和S-Sepharose层析，以21%的收率从经肝毒素处理的小鼠肝脏上清液中纯化得到，纯度约28万倍。非还原条件下纯化的mHGF sds -聚丙烯酰胺凝胶电泳（PAGE）显示为单个分子量为76K的条带，还原后的mHGF SDS-PAGE检测到两个分子量为62K和31K的条带，表明mHGF为异源二聚体。mHGF对培养肝细胞DNA合成的刺激作用与hHGF一样有效。mHGF的作用与EGF和酸性FGF的最大作用是叠加的，与胰岛素的最大作用是协同的，而与hHGF的最大作用没有协同作用。因此，mHGF的性质与hHGF相似。mHGF可能是一种与hHGF结构不同的hHGF样因子，其活性不受兔抗hHGF抗血清的抑制，而兔抗hHGF抗血清能中和hHGF活性。