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Immunomodulating Effect of Extract of Cheonggukjang Fermented with Bacillus amyloliquefaciens (SRCM100730) on RAW 264.7 Macrophages

Bacillus amyloliquefaciens(SRCM 100730)로 발효된 청국장 추출물의 RAW 264.7 대식세포 면역증강 활성

  • Choo, Seung Bin (Department of Microbiology, College of Medicine, Konyang University) ;
  • Yang, Hui (Department of Microbiology, College of Medicine, Konyang University) ;
  • Jeong, Do-Yuon (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Jeong, Seong-Yeop (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Ryu, Myeong Seon (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Oh, Kwang-Hoon (Department of Physical Education, College of Education, Kongju National University) ;
  • Yoo, Yung Choon (Department of Microbiology, College of Medicine, Konyang University)
  • 추승빈 (건양대학교 의학과 미생물학교실) ;
  • 양혜 (건양대학교 의학과 미생물학교실) ;
  • 정도연 ((재)발효미생물산업진흥원) ;
  • 정성엽 ((재)발효미생물산업진흥원) ;
  • 류명선 ((재)발효미생물산업진흥원) ;
  • 오광훈 (공주대학교 사범대학 체육교육과) ;
  • 유영춘 (건양대학교 의학과 미생물학교실)
  • Received : 2017.09.05
  • Accepted : 2017.11.03
  • Published : 2017.11.30

Abstract

Cheonggukjang is well known as a traditional fermented food in Korea and has various biological activity. In this study, immune-enhancing activity of extract of cheonggukjang fermented with Bacillus amyloliquefaciens (SRCM100730) was examined in RAW 264.7 murine macrophages. Treatment with extract augmented production of nitric oxide (NO) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) from RAW 264.7 macrophages in a dose-dependent manner. Similarly, increased mRNA expression of inducible nitric oxide synthase (iNOS) and $TNF-{\alpha}$ was observed. In addition, the extract synergistically enhanced production of NO and $TNF-{\alpha}$ from lipopolysaccharide (LPS)-stimulated macrophages. Analysis of intracellular pathways revealed that the immune-enhancing activity of cheonggukjang extract was related to activation of mitogen-activated protein kinases (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). These results suggest that cheonggukjang fermented with B. amyloliquefaciens (SRCM100730) is a beneficial food effective for activation of immune responses.

청국장은 한국의 전통발효식품으로서 다양한 생리활성이 보고되어 있다. 본 연구에서는 B. amyloliquefaciens 균종에 속하는 신규한 균주인 B. amyloliquefaciens(SRCM 100730)에 의한 발효 청국장이 선천면역 세포인 RAW 264.7 대식세포를 활성화하는 효과를 가지는가를 검토하였다. 청국장 추출물을 RAW 264.7 세포에 처리한 결과 처리농도에 의존하여 $TNF-{\alpha}$와 NO의 생성이 증가하였으며, 이러한 면역조절 물질의 증가는 iNOS와 $TNF-{\alpha}$ mRNA 발현에서도 확인되었다. 또한, 청국장 추출물에 의한 대식세포 활성화와 관련한 세포 내 작용기전을 해석한 결과 청국장 처리에 의해 p38과 ERK와 같은 MAPK의 인산화와 $NF-{\kappa}B$ 활성화가 유도되는 것으로 밝혀졌다. 하지만 MAPK에 속하는 JNK에 대해서는 아무런 영향을 주지 않는 것으로 나타났다. 한편 청국장 추출물을 LPS와 함께 처리한 실험에 있어서 청국장 추출물은 LPS에 의한 대식세포 활성화를 촉진하여 $TNF-{\alpha}$와 NO의 생성을 증가시키는 시너지 효과를 나타내었다. 이들 결과로부터 B. amyloliquefaciens(SRCM 100730) 균주로 발효한 청국장 추출물은 대식세포를 활성화하여 선천면역을 증가시키는 효과가 있는 것으로 확인되었다.

Keywords

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