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추출조건에 따른 차가버섯 생리활성 및 면역활성 다당

Pharmacological Activity of Chaga Mushroom on Extraction Conditions and Immunostimulating Polysaccharide

초록

추출조건에 따른 생리활성 영향을 검토하기 위하여 차가버섯(Inonotus obliquus)을 물($50^{\circ}C$ reflux, $90^{\circ}C$ 이상 decoction, $12^{\circ}C$ 가압추출)과 에탄올(50, 70과 $90^{\circ}C$ reflux)로 추출한 후 물 추출물을 조다당획분(IO-CP)으로 분획하였을 때 IO-CP(4.8~16.8%)의 수율은 에탄올추출물(IO-E, 1.9~2.7%)보다 높았으며 추출온도 증가에 따라 그 차이는 더 컸다. 가압추출로 조제된 조다당획분(IO-CP-121)은 가장 높은 총 폴리페놀 및 플라보노이드 함량(35.10 mg TAE/g와 18.48 mg QE/g)과 DPPH와 ABTS를 이용한 자유 라디칼소거능(26.08과 27.99 mg AEAC/100 mg)을 나타내었다. IO-CP-D(decoction)와 IO-CP-50(reflux)은 IO-CP-121보다 유의적으로 높은 마이토젠(saline 대조군의 2.10과 1.95배, 100 ${\mu}g/mL$)과 장관면역 활성(6.30과 5.74배)을 보였으나 에탄올추출물 활성은 확인되지 않았다. 또한, 모든 IO-CP는 0.1 mg/mL 농도에서 RAW 264.7 세포주에 대하여 독성을 나타내지 않았으나, IO-CP-121이 LPS-자극 RAW 264.7 세포주에 대하여 가장 유의적으로 염증성 인자인 TNF-${\alpha}$와 nitric oxide(NO) 생성을 억제하였다(29.2와 63.5%). 에탄올추출물도 0.1 mg/mL 농도에서 독성을 보이지 않았으나 TNF-${\alpha}$와 NO의 생성 억제능은 IO-CP-121보다 현저히 낮음을 알수 있었다. 한편, 활성 다당획분을 분리하기 위하여 면역활성 획분인 IO-CP-D를 DEAE-Sepharose CL-6B column에서 비흡착 획분(IO-CP-I)과 7개의 흡착 획분(IO-CP-II~VIII)으로 분획한 결과, IO-CP-II가 가장 우수한 마이토젠과 마크로파지 활성(4.51과 1.64배)을 나타내었다. 이와 같이 분리된 면역활성 획분인 IO-CP-II는 주로 중성당(61.86%)과 함께 소량의 산성당(2.96%)을 포함하고 있는 다당류임이 밝혀졌으며, 주요 구성당으로서 Glc, Gal와 Man(molar ratio of 1.00:0.55:0.31)를 포함하고 있음을 확인할 수 있었다. 이러한 결과로부터 추출조건은 차가버섯 생리활성에 중요하게 작용하고 있으며, decoction 추출방법으로 제조하여 분획한 면역활성 획분으로부터 차가버섯 면역활성은 적어도 중성 다당류가 관여하고 있음을 알 수 있었다.

Abstract

To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

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이 논문을 인용한 문헌 (1)

  1. Oh, Seong-Yoon ; Chung, Mi Ja ; Choi, Jae-Ho ; Oh, Deog-Hwan 2014. "Screening of Personalized Immunostimulatory Activities of Saengsik Materials and Products Using Human Primary Immune Cell" 한국식품영양과학회지 = Journal of the Korean Society of Food Science and Nutrition, 43(9): 1325~1333 

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