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Antitumor and Immuno-potentiating Activities of Crude Polysaccharides from Fruiting Body of Agaricus brasiliensis

신령버섯(Agaricus brasiliensis) 자실체 추출 조다당류의 항암 및 면역증강 작용

  • Received : 2011.03.15
  • Accepted : 2011.03.21
  • Published : 2011.04.01

Abstract

Agaricus brasiliensis, one of edible mushroom belonging to Basidiomycota, has been used for curing gastric ulcer and stomach cancer of human beings and also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were prepared from fruiting body of the mushroom. ${\beta}$-glucan and total protein contents were identify from fractions of edible mushrooms extract. The ${\beta}$-glucan and protein contents of all fractions of the mushrooms ranged from 21.54~32.31% and 0.16~9.34%, respectively. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, NIH3T3 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 18.8~50.6% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cell by 1.2 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. MeOH and Na improved the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.6 fold compared with control at the concentration of 50~500 ${\mu}g/ml$. Fr. Na generated 15.9 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7 at the concentration of 200 ${\mu}g/ml$, while lipopolysaccharide, a positive control, produced 3.7 ${\mu}M$. The Fr. NaCl, Fr. HW and Fr. MeOH increased the secretion of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by 2.2 times compared with the control group. Fr. Na increased the numbers of peritoneal exudate cells by 4 folds at the concentration of 50mg/kg compared with control. Circulating leukocytes increased by 2.7 folds when Fr. HW from A. brasiliensis was inoculated at the concentration of 50 mg/kg body weight. The hematological and blood chemical analysis of the 3 fractions did not show any difference in blood compositions and enzyme activities compared with the control group (p<0.05). Therefore, the experimental results suggested that crude polysaccharides extracted from A. brasiliensis contain antitumor and immuno-potentiating activities against Sarcoma 180 in ICR mice.

신령버섯은 담자균문, 주름버섯목, 주름버섯과에 속하는 식용버섯으로 혈당, 혈압강하효과와 콜레스테롤 저하, 항종양, 암예방 효과가 있는 것으로 알려져 있다. 신령버섯의 자실체로부터 메탄올, 중성염용액 및 열수 등을 이용하여 조다당류를 추출하고 성분을 분석한 결과 ${\beta}$-glucan은 21.54~32.31%, 단백질은 0.16~9.34%로 구성되어 있는 것이 밝혀졌고 신령버섯 추출 조다당류를 Sarcoma 180, HT-29, NIH3T3 및 RAW 264.7 등의 암세포 및 정상세포에 대한 독성을 조사한 결과 10~2000 ${\mu}g/ml$의 조다당류 농도에서 각각의 세포는 세포독성을 나타내지 않았다. Sarcoma 180으로 접종된 ICR 생쥐에 신령버섯의 자실체에서 추출한 각각의 조다당류를 투여한 실험군은 대조군에 비해 수명이 각각 18.8~50.6% 연장되었다. 비장세포의 증식능과 B 임파구의 활성화에 미치는 alkaline phosphatase의 활성을 조사한 결과 비장 세포 증식능은 대조군에 대해 1.1~1.2배의 증식능을 보였고 B 임파구의 활성은 대조군에 대해 1.2~1.6배 증가하였다. 또한 버섯추출 조다당류를 투여한 실험군이 대조군에 비하여 대식세포에서 1.3~4.3배의 많은 nitric oxide를 발생시켰다. 비장세포에 여러 농도의 조다당류를 처리한 후 TNF-${\alpha}$, IL-$1{\beta}$, IL-2 및 IL-6 등의 사이토카인 분비량을 측정한 결과 생성된 사이토카인의 양은 대조군에 비해 2.2배 높게 나타났다. 중성염용액추출 조다당류를 50 mg/kg body weight의 농도로 투여한 실험군 생쥐의 총 복강 세포 수는 대조군에 비하여 4배 증가하였고, 열수추출 조다당류를 50 mg/kg body weight의 농도로 투여한 실험군 생쥐의 백혈구 수는 대조군에 비하여 각각 2.7배 증가하였다. 혈액생화학적 검사를 시행한 결과, 대조군과 유사한 경향을 나타냈다. 따라서 신령버섯의 자실체에서 추출한 조다당류는 생쥐의 면역을 증강시키는 것은 물론 Sarcoma 180에 대한 항암효과를 나타내는 것으로 밝혀졌다.

Keywords

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