Identification of core active disaccharides in heparin for HGF-inducing activity.
Identification of core active disaccharides in heparin for HGF-inducing activity.
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DOI：
10.4103/0976-500x.155483
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发表时间：
2015-04
影响因子：
0.2
通讯作者：
Nakamura T
中科院分区：
文献类型：
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作者：
Kato T;Sakiyama R;Oka K;Nakamura T
为确定不同硫酸化肝素二糖诱导肝细胞生长因子(HGF)活性的基团位置，探讨肝素二糖体内是否升高HGF水平，从而对急性肝损伤起到保护作用。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)-GlcNAc(6S)加入MRC-9成纤维细胞，用酶联免疫吸附分析法检测培养液中肝细胞生长因子的浓度。给C57BL/6小鼠注射ΔUA-GlcNS(10 0μg/头)，12h后测定小鼠血浆肝细胞生长因子水平，用15 mg/kg四氯化碳诱导小鼠急性肝炎后，取肝标本行苏木精-伊红染色。在所研究的双糖中，ΔUA-GlcNS、ΔUA-GlcNAc(6S)和ΔUA-GlcNS(6S)能刺激成纤维细胞产生肝细胞生长因子。然而，尽管存在N-硫化和/或6-O-硫化双糖，2-O-硫化双糖[ΔUA(2S)-GlcNS，ΔUA(2S)-GlcNAc(6S)和ΔUA(2S)-GlcNS(6S)]都没有显示任何活性。由此可见，己糖醛酸的2-O-磺化反应具有抑制作用。此外，ΔUA-GlcNS可提高正常小鼠血浆中肝细胞生长因子的水平，并能预防四氯化碳所致的小鼠肝损伤。氨基葡萄糖与非硫化己糖酸的N-硫化和/或6-O-硫化反应为二糖的HGF诱导活性提供了结构基础。ΔUA-GlcNS可提高血浆中肝细胞生长因子的水平，对四氯化碳所致的急性肝损伤具有保护作用。
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.