Stimulation of hepatocyte growth factor production by heparin-derived oligosaccharides.
We previously reported that heparin post-transcriptionally stimulates the production of hepatocyte growth factor (HGF). In this study, we addressed the size-dependency of heparin fragments on the HGF-inducing activity aiming to obtain fragments without antiblood coagulant activity. Heparin fragments, produced by digestion with heparinase, were size-fractionated and tested for HGF-inducing activity in cultured human fibroblasts. The HGF-inducing activity deceased with the reduction in oligosaccharide size. Decasaccharides exerted an activity comparable with undigested heparin, while smaller oligosaccharides showed lesser activities. The anticoagulant activity of heparin fragments also decreased with size and anticoagulant activity of decasaccharides was <13% that of undigested heparin. Further fractionation of decasaccharides by anion-exchange chromatography revealed that most of the decasaccharides had HGF-inducing activity and the extent of sulfation was roughly related to the activity. The lack of N-sulfation in heparin markedly reduced HGF-inducing activity, whereas 2-O-desulfation or 6-O-desulation had a lesser influence. Moreover, an N-sulfated disaccharide showed significant HGF-inducing activity, suggesting the involvement of N-sulfation in HGF-inducing activity. Because of the much reduced anticoagulant activity, potential applications of heparin-derived oligosaccharides such as decasaccharides is considerable as a therapeutic agent for many diseases.
Sakiyama
Ryoichi
R
Division of Molecular Regenerative Medicine, Department of Biochemistry and Molecular Biology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
Fukuta
Kazuhiro
K
Matsumoto
Kunio
K
Furukawa
Masumi
M
Takahashi
Yoshihiro
Y
Nakamura
Toshikazu
T
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D017077:Culture Media, Conditioned; D006493:Heparin / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis*; D006801:Humans; D009844:Oligosaccharides / Q000494:pharmacology*; D034421:Sus scrofa
D000925:Anticoagulants; D017077:Culture Media, Conditioned; D009844:Oligosaccharides; D017228:Hepatocyte Growth Factor; D006493:Heparin