Masumoto A, Yamamoto N
Department of Functional Morphology, School of Nursing Kitasato University, Kanagawa.
Cell Struct Funct. 1993 Apr;18(2):87-94. doi: 10.1247/csf.18.87.
A growth factor stimulating DNA synthesis of adult rat hepatocytes in primary culture was found in the conditioned medium after culturing nonparenchymal liver cells (NPC). Adding heparin to the NPC cultured medium stimulated the growth-factor secretion from NPC. The growth factor was secreted mainly by Kupffer cells. The partially purified growth factor from the NPC appeared to be related to the HGF isolated from platelets according to three criteria: (a) binding to Heparin-Sepharose and eluting at about 0.65 M NaCl, (b) having a Mr of about 70 kDa, (c) having an immunoreactivity to antibody against rat platelet-derived HGF. Adding heparin to the NPC cultured medium also resulted in protection of the growth factor from heat- and acid-inactivation, but the direct interaction of heparin with the partially purified growth factor did not lead to such protection. Perfusion of normal adult rat livers with Hanks' solution containing 1 M NaCl in situ led to the release of large amounts of hepatocyte growth factor (10). These findings suggest that the hepatocyte growth factor derived from NPC binds to ECM between the layer of hepatocytes and endothelial cells forming the sinusoids in normal adult rat liver and that this may play a role in stabilization and maintaining the pool of HGF, which functions to constantly supply HGF in the setting of liver regeneration.
在对非实质肝细胞（NPC）进行培养后，在其条件培养基中发现了一种能刺激原代培养的成年大鼠肝细胞DNA合成的生长因子。向NPC培养基中添加肝素可刺激NPC分泌生长因子。该生长因子主要由库普弗细胞分泌。根据三个标准，从NPC中部分纯化得到的生长因子似乎与从血小板中分离出的肝细胞生长因子（HGF）相关：（a）与肝素-琼脂糖结合并在约0.65M NaCl浓度下洗脱；（b）分子量约为70kDa；（c）对大鼠血小板衍生的HGF抗体具有免疫反应性。向NPC培养基中添加肝素还能使生长因子免受热和酸的灭活作用，但肝素与部分纯化的生长因子直接相互作用并不会产生这种保护作用。用含1M NaCl的汉克斯溶液原位灌注正常成年大鼠肝脏会导致大量肝细胞生长因子的释放。这些发现表明，源自NPC的肝细胞生长因子与正常成年大鼠肝脏中形成肝血窦的肝细胞层和内皮细胞层之间的细胞外基质（ECM）结合，这可能在稳定和维持HGF池中发挥作用，而HGF池在肝脏再生过程中持续供应HGF。
