Isolation and Characterization of Immunomodulatory Glycoprotein from the Root of Panax ginseng

  • Published : 2000.09.01

Abstract

A high molecular (more than 10 kDa) fraction, showing mitogenic and comitogenic activities in spleen cells of mouse, was isolated from water extract of ginseng. The crude protein substance prepared by 80% (NH$_4$)$_2$SO$_4$ precipitation from this fraction was purified and isolated by DEAE Sepharose column chromatography. Among the fractions eluted, it was found that four kinds of fractions eluted with 0 to 1 M NaCl gradient were glycoproteins, which induced proliferation of spleen cells and increased NO production in macrophages. Among them, F-2 fraction, which contained 35.9% protein,49.4% neutral sugar and 12.5% uronic acid, was found to show mitogenic activity as strong as that of LPS (lipopolysaccharide) at a concentration of 100 $\mu\textrm{g}$/ml and to remarkably stimulate NO production by murine macrophages at a concentration of 500 $\mu\textrm{g}$/ml. When F-2 is deproteinized, the mitogenic activity of F-2 was decreased significantly to 70.9% as compared with that of F-2. This results suggests that the protein moiety of F-2 may play an important role in immunomodulating activity of glycoprotein from the root of Panax ginseng.

인삼(Panaxginseng)의 물추출물로부터 분리한 분자량 10 kDa 이상의 분획에서 강한 비장세포 분열증식 효과를 볼 수 있었다. 또한 이 분획은 LPS와 Con A에 의해서 유도된 B-cell과 T-cell의 증식효과를 66%와 49%씩 더 증가시켰다. 이 분획을 80% ammonium sulfate 침전법을 실시해서 조 단백질 분획을 얻었으며 이를 다시 DEAE Sepharose CL-6B column chromatography를 수행해서 단백질 분획들을 분리하였다. 대부분의 단백질은NaCl gradien에 의해서 용출 되었고 0~1 M NaCl gradient에 의해서 분리된 4종류의 당단백질 분획에서 비장세포 증식효과와 대식세포의 NO 생산촉진 효과가 나타났다. 특히 단백질 35.9 %,중성당 49.4%, 산성당 12.5%로 구성되어있는 F-2 분획에서는 LPS수준의 높은 면역세포 증식효과를 보였다. 그러나 F-2를 protease로 처리한 후의 비장세포 증식 효과는 대조군과 대비해서 29.1%가 낮아졌다. 이상의 결과로부터 인삼의 물추출물에서 분리된 F-2분획의 면역세포 활성효과에는 단백질 부분이 중요한 역할을 한다고 생각된다.

Keywords