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Selective depletion of glycyrrhizin from Si-Ni-San, a traditional Chinese prescription, blocks its effect on contact sensitivity in mice and recovers adhesion and metalloproteinases production of T lymphocytes.

Abstract
In the present study, we performed to selectively deplete glycyrrhizin from Si-Ni-San, a traditional Chinese prescription that consists of 4 Chinese herbs including Radix Glycyrrhizae Uralensis, and examined its influence on the suppressing activity of Si-Ni-San against contact sensitivity in mice. An immunoaffinity column was made by covalently coupling the polyclonal antibody, obtained by the immunization with glycyrrhizin-BSA conjugate, to CNBr-activated Sepharose 4B. By using this column, glycyrrhizin in Si-Ni-San was selectively and almost completely depleted from the whole extract, which was confirmed by high-performance liquid chromatography (HPLC). Both 200 mg/kg of Si-Ni-San and 10 mg/kg of glycyrrhizin, the dose corresponding to its proportion contained in Si-Ni-San, significantly reduced the ear swelling of picryl chloride (PCl)-induced ear contact sensitivity in mice and the inhibition by Si-Ni-San was stronger than that by glycyrrhizin. The adhesion activity to type IV collagen of the isolated spleen cells from PCl-sensitized mice was significantly decreased by both Si-Ni-San and glycyrrhizin. However, the glycyrrhizin-depleted sample of Si-Ni-San (Si-Ni-San(GL-)) only showed a slight inhibition on the cell adhesion. Furthermore, the spleen cells from PCl-sensitized mice produced more matrix metalloproteinase (MMP)-2 and -9 than naive spleen cells did, and both Si-Ni-San and glycyrrhizin remarkably reduced MMP-2 and MMP-9 production. In contrast, Si-Ni-San(GL-) only showed a slight inhibition. These results suggest that glycyrrhizin may act as one of the active constituents of Si-Ni-San in inhibiting delayed-type hypersensitivity reaction via down-regulating the MMP production and the cell adhesion to extracellular matrix. The present study also provides a new approach to recognize and validate an active constituent in traditional prescription through a selective depletion.
AuthorsLi Zhang, Yang Sun, Ting Chen, Qiang Xu
JournalInternational immunopharmacology (Int Immunopharmacol) Vol. 5 Issue 7-8 Pg. 1193-204 (Jul 2005) ISSN: 1567-5769 [Print] Netherlands
PMID15914324 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Glycyrrhizic Acid
  • Dexamethasone
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9
Topics
  • Animals
  • Cell Adhesion (drug effects)
  • Chromatography, Affinity
  • Dermatitis, Contact (drug therapy)
  • Dexamethasone (pharmacology)
  • Female
  • Glycyrrhizic Acid (immunology, therapeutic use)
  • Matrix Metalloproteinase 2 (biosynthesis)
  • Matrix Metalloproteinase 9 (biosynthesis)
  • Medicine, Chinese Traditional
  • Mice
  • Mice, Inbred ICR
  • T-Lymphocytes (drug effects, enzymology)

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