Kato Takashi, Sakiyama Ryoichi, Oka Kiyomasa, Nakamura Toshikazu
Department of Pathology, Faculty of Medicine, Kinki University, 377-2 Ohno-Higashi, Osaka Sayama, Osaka, 565-0871, Japan ; Department of Clinical Engineering, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, Japan.
Research and Development, Neurogen Inc., 7-7-20 Saito-asagi, Ibaraki, Osaka 567-0085, Japan.
J Pharmacol Pharmacother. 2015 Apr-Jun;6(2):77-82. doi: 10.4103/0976-500X.155483.
To ascertain the positions of sulfated groups for HGF-inducing activity using differently sulfated heparin disaccharides and to investigate whether the heparin disaccharide elevates HGF levels in plasma in vivo and exerts protective effects on acute liver injury.
The heparin disaccharides ΔUA-GlcNS, ΔUA (2S)-GlcN, ΔUA-GlcNAc (6S), ΔUA-GlcNS (6S), ΔUA (2S)-GlcNS, ΔUA (2S)-GlcNAc (6S), ΔUA-GlcNAc and ΔUA (2S)-GlcNS (6S) were added to MRC-9 fibroblasts and HGF concentrations in culture media were determined by enzyme-linked immunosorbent assay. Furthermore, ΔUA-GlcNS (100 μg/head) was injected into C57BL/6 mice and plasma levels of HGF measured at 12 h. After acute hepatitis was induced by CCl4 (15 mg/kg) in mice, liver specimens were stained with hematoxylin and eosin (H and E). Levels of aspartate aminotransferase and alanine aminotransferase were measured at 24 h.
Among the disaccharides investigated, ΔUA-GlcNS, ΔUA-GlcNAc (6S) and ΔUA-GlcNS (6S) stimulated HGF production in MRC-9 fibroblasts. However, none of the 2-O-sulfated disaccharides [ΔUA (2S)-GlcNS, ΔUA (2S)-GlcNAc (6S) and ΔUA (2S)-GlcNS (6S)] showed any activity despite the presence of N-sulfated and/or 6-O-sulfated disaccharides. Thus, 2-O-sulfation of hexuronic acid has an inhibitory effect. Moreover, ΔUA-GlcNS administration increased plasma levels of HGF in normal mice and prevented CCl4-induced liver injury in mice.
N-sulfation and/or 6-O-sulfation of glucosamine with nonsulfated hexuronic acid provides a structural basis for the HGF-inducing activity of disaccharides. ΔUA-GlcNS increases plasma levels of HGF and protects against CCl4-induced acute liver injury.
使用不同硫酸化的肝素二糖确定硫酸化基团对肝细胞生长因子（HGF）诱导活性的位置，并研究肝素二糖是否能在体内提高血浆中HGF水平以及对急性肝损伤发挥保护作用。
将肝素二糖ΔUA-GlcNS、ΔUA(2S)-GlcN、ΔUA-GlcNAc(6S)、ΔUA-GlcNS(6S)、ΔUA(2S)-GlcNS、ΔUA(2S)-GlcNAc(6S)、ΔUA-GlcNAc和ΔUA(2S)-GlcNS(6S)添加到MRC-9成纤维细胞中，通过酶联免疫吸附测定法测定培养基中的HGF浓度。此外，将ΔUA-GlcNS（100μg/只）注射到C57BL/6小鼠体内，并在12小时时测量血浆中HGF水平。在用四氯化碳（15mg/kg）诱导小鼠急性肝炎后，将肝脏标本用苏木精和伊红（H&E）染色。在24小时时测量天冬氨酸转氨酶和丙氨酸转氨酶水平。
在所研究的二糖中，ΔUA-GlcNS、ΔUA-GlcNAc(6S)和ΔUA-GlcNS(6S)刺激MRC-9成纤维细胞产生HGF。然而，尽管存在N-硫酸化和/或6-O-硫酸化的二糖，但所有2-O-硫酸化的二糖[ΔUA(2S)-GlcNS、ΔUA(2S)-GlcNAc(6S)和ΔUA(2S)-GlcNS(6S)]均未显示出任何活性。因此，己糖醛酸的2-O-硫酸化具有抑制作用。此外，给予ΔUA-GlcNS可提高正常小鼠血浆中HGF水平，并预防小鼠四氯化碳诱导的肝损伤。
氨基葡萄糖的N-硫酸化和/或6-O-硫酸化与非硫酸化的己糖醛酸为二糖的HGF诱导活性提供了结构基础。ΔUA-GlcNS可提高血浆中HGF水平，并预防四氯化碳诱导的急性肝损伤。
