American Society of Hirudotherapy

Characterisation of separated end hyaluronan oligosaccharides from leech hyaluronidase and evaluation of angiogenesis

Research article published in Carbohydr Polym (2016)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Preclinical (animal)Salivary PharmacologyDrug DevelopmentLv M et al. · Carbohydr Polym, 2016

Abstract

Hyaluronan oligosaccharides (o-HAs), especially saturated o-HAs, have attracted intensive attention due to their potential applications in medical treatments. In this study, the hydrolysis process of leech hyaluronidase (LHase) towards the hyaluronan was investigated by HPLC and HPLC/ESI-MS. The proportions of hyaluronan tetrasaccharide (HA4) with hexasaccharide (HA6), end products, were illustrated to have a relationship with the amount of LHase. Higher yield of HA4 was achieved with higher activity of LHase. After optimisation of the packing resin and operation parameters (balanced pH, elution concentration, elution volume and elution flow rate), the highly pure HA4 and HA6 were efficiently separated and prepared by combining ion exchange Q-Sepharose Fast Flow and size exclusion column chromatography. Compared with o-HAs (average Mr of 4000 Da), HA4 and HA6 were demonstrated to show higher activity for promoting angiogenesis, which was similar with the corresponding HA4 and HA6 produced by bovine testicular hyaluronidase. The pure HA4 and HA6 that prepared from LHase will attract intensive studies and be used in potential applications in near future.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal ArticleResearch Support, Non-U.S. Gov't
Indexed MeSH termsAngiogenesis Inducing AgentsAnimalsChromatography, GelChromatography, High Pressure LiquidFemaleHyaluronic AcidHyaluronoglucosaminidaseHydrolysisLeechesMiceNeovascularization, PhysiologicOligosaccharides

Summary

Pure HA4 and HA6 oligosaccharides separated from leech hyaluronidase hydrolysate; show higher angiogenic activity than larger oligosaccharides.

Why This Matters for Hirudotherapy

This study investigated the hydrolysis of hyaluronan by leech hyaluronidase (LHase), characterizing end products by HPLC and HPLC/ESI-MS, and optimized separation of highly pure tetrasaccharide (HA4) and hexasaccharide (HA6) via ion-exchange and size-exclusion chromatography. HA4 and HA6 demonstrated higher angiogenesis-promoting activity compared to larger oligosaccharides, similar to corresponding products from bovine testicular hyaluronidase. This is relevant to ASH as it examines enzymatic activity of a component derived from leeches and its bioactive oligosaccharide products. CAVEAT: The abstract does not specify the experimental system used for the angiogenesis assessment, and no human therapeutic application or hirudotherapy is described; the medical relevance noted is prospective, as stated in the abstract.

Citation

Characterisation of separated end hyaluronan oligosaccharides from leech hyaluronidase and evaluation of angiogenesis.

Lv M et al. · Carbohydr Polym, 2016

DOI

Added to ASH library: May 27, 2026 · Site last updated: June 18, 2026

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