Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Immunomodulatory and anti-metastatic activities of a crude polysaccharide isolated from Korean apple vinegar

한국산 사과식초에서 분리한 다당의 면역 및 항전이 활성

  • Kim, Han Wool (Department of Food Science and Biotechnology, Kyonggi University) ;
  • Shin, Kwang-Soon (Department of Food Science and Biotechnology, Kyonggi University)
  • 김한울 (경기대학교 식품생물공학과) ;
  • 신광순 (경기대학교 식품생물공학과)
  • Received : 2019.01.14
  • Accepted : 2019.01.31
  • Published : 2019.04.30
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Abstract

To characterize new physiologically active components in Korean apple vinegar, a crude polysaccharide (KAV-0) was prepared by precipitation with 80% (v/v) ethanol. KAV-0 mainly comprises 38.2% mannose, 19.1% galactose and 14.3% glucose. In an in vitro cytotoxicity analysis, KAV-0 promoted the proliferation of peritoneal macrophages and RAW 264.7 cells in a dose-dependent manner, and showed no cytotoxicity in B16-BL6 melanoma cells. Murine peritoneal macrophages and RAW 264.7 cells stimulated by KAV-0 produced various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide (NO). Intravenous (i.v.) administration of KAV-0 significantly augmented NK cell cytotoxicity against Yac-1 tumor cells. In experimental lung metastasis caused by B16-BL6 melanomas, prophylactic i.v. administration of KAV-0 at a dosage of $1,000{\mu}g/mouse$ inhibited lung metastasis by 53.0%. These results suggest that the crude polysaccharide (KAV-0) isolated from Korean apple vinegar has a considerably high anti-metastatic activity and immunomodulatory activities beneficial to human health.

전통발효식초에 속하는 사과식초로부터 에탄올 및 증류수를 이용, 다당체를 분리하여 새로운 면역활성 물질을 확인하고자 하였다. 사과식초를 농축하고, 80% 에탄올을 가하여 침전물을 얻었으며 소량에 물에 용해하여 투석 및 동결건조를 거쳐 조다당 KAV-0를 얻었다. 구성당 분석 결과, KAV-0는 주로 mannose 38.2%, galactose 19.1% 및 glucose 14.3%로 구성되어있음을 확인하였고, 정상세포주(RAW 264.7 세포 및 대식세포) 및 종양세포주(B16-BL6 melanoma 세포)에 대해 세포독성 측정 결과, 직접적인 세포독성을 나타내지 않았다. 대식세포 유래 세포주인 RAW 264.7 세포 자극에 대한 사이토카인 및 산화질소 생산능을 측정한 결과, IL-6, $TNF-{\alpha}$ 및 산화질소 모두 농도 의존적으로 분비능이 증가하고 저농도에서도 우수한 활성을 나타냈으며, 마우스 복강유래 대식세포 자극에 대한 사이토카인 및 산화질소 생산능에서도 동일한 양상의 결과를 보여 사과식초 유래 조다당 KAV-0가 높은 대식세포 활성능을 소유함을 확인할 수 있었다. 한편, KAV-0의 정맥투여에 따른 NK 세포 활성화를 측정한 결과(ex vivo), 시료에 의해 활성화된 NK 세포는 Yac-1 종양 세포주에 대한 치사활성을 농도 의존적으로 증진시켰으며, $1,000{\mu}g/mouse$에서 월등히 우수한 NK 세포 활성능을 유도하였다. 또한, B16-BL6 melanoma를 이용한 폐암전이모델에서 KAV-0를 정맥투여한 뒤 항전이 활성을 측정한 결과, 농도 의존적으로 암 저해 효과가 나타났으며, 특히 $1,000{\mu}g/mouse$에서는 약 53%의 우수한 암전이 억제 활성을 나타냈다. 이상의 결과로부터 사과식초 유래 조다당 KAV-0는 인체의 면역계를 자극하여 우수한 면역활성 및 항전이 효과를 나타낼 수 있음이 확인되었고 건강 기능성 소재로의 활용 가능성이 충분할 것으로 최종 판단되었다.

Keywords

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Fig. 1. Isolation of (immuno-stimulating) polysaccharide (KAV- 0) isolated from Korean apple vinegars (A), and elution pattern and MW-determination of KAV-0 on HPSEC (B).

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Fig. 2. Effect of KAV-0 on viabilities of murine peritoneal macrophages, RAW 264.7 and B16-BL6 melanoma cells.

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Fig. 3. Effect of KAV-0 on cytokines and nitric oxide production by RAW 264.7 cells. RAW 264.7 cells were treated with indicated concentrations of KAV-0 for 24 hr.

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Fig. 4. Effect of KAV-0 on cytokines and nitric oxide production by murine peritoneal macrophage in vitro.

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Fig. 5. Effect of KAV-0 on cytolytic activity of NK cells ex vivo.

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Fig. 6. Inhibitory effect of KAV-0 on lung metastasis produced by i.v. inoculation of B16-BL6 melanoma cells.

Table. 1. Chemical properties of KAV-0 isolated from apple vinegar

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