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천식 마우스 모델의 알러지 반응에서 Enterococcus faecalis 전체 추출물의 억제 효과

The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model

  • 장정현 (대구한의대학교 임상병리학과) ;
  • 양은주 (대구한의대학교 임상병리학과) ;
  • 유선녕 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 안순철 (부산대학교 의학전문대학원 미생물학 및 면역학 교실)
  • Chang, Jeong Hyun (Department of Clinical Laboratory Science, Daegu Haany University) ;
  • Yang, EunJu (Department of Clinical Laboratory Science, Daegu Haany University) ;
  • Yu, Sun Nyoung (Department of Microbiology and Immunology, Pusan National University School of Medicine) ;
  • Ahn, Soon-Cheol (Department of Microbiology and Immunology, Pusan National University School of Medicine)
  • 투고 : 2017.08.24
  • 심사 : 2017.10.25
  • 발행 : 2017.10.30

초록

프로바이오틱스(probiotics)는 숙주에 이로운 작용을 하는 장내세균으로 주로 알려져 있으며 꾸준히 염증치료의 새로운 치료물질로 주목 받고 있다. 그러나 Lactobacillus나 Bifidobacterium과 같은 전형적인 프로바이오틱스는 장내에서 꾸준히 생존하기 어렵고 그로 인해 매개할 수 있는 다양한 효능이 줄어든다. 반면, Enterococcus는 다양한 물질을 분비하고 생물학적 활성을 지니고 있는 프로바이오틱스이긴 하나 동시에 기회감염균으로 항생제 내성능을 쉽게 획득할 수도 있어 사용을 자제할 것을 권고하고 있으며, Vancomycin 내성인자를 원래부터 가지고 있어서 이 인자의 발현과 함께 타 독성균주에 전달할 가능성이 있다. 그러나 본 연구에서는 이러한 Enterococcus의 추출물을 이용하여 항염증 물질을 확인해보고자 하였다. 특히, 호흡기계 질환인 천식의 치료효능을 보기 위해 human monocytic cell line인 THP-1 세포를 이용한 in vitro test와 천식 유발 BALB/c mice를 이용한 in vivo test를 시행하였다. 천식은 사이토카인과 IgE 등 세포 염증성 물질을 분비하고 호산구, 호중구, 호염기구, 비만세포, 단구, T 세포 등 다양한 염증 세포가 폐 부위로 침윤되는 복합적인 염증 질환으로 잘 알려져 있다. 본 연구에서는 우선 Enterococcus faecalis 에서 초음파 처리를 통해 균체성분을 분해한 다음 동결건조하여 추출물을 획득하였다. E. faecalis에서 추출한 추출물(Ef extract)을 PBS에 부유시킨 다음, 염증유발을 위해 ovalbumin (OVA)를 처리한 THP-1 세포에 처리한 결과, OVA에 의해 증가한 THP-1 세포의 생존율이 감소하였다. 또한 OVA에 의해 천식이 유발된 BALB/c mice에서 염증부위인 폐의 세척액을 채취하여 폐 조직 내 세포 변화를 관찰하였다. 그 결과, Ef extract에 의해 호산구와 호중구가 Ef extract 농도에 따라 점차 감소하였고 혈액 내에서도 Ef extract에 의해 호산구, 호중구, 호염기구가 감소하였다. 이는 Ef extract에 의해 염증세포로부터 IL-5의 분비를 억제하고 IgE의 양을 감소시켜 이와 관련된 염증세포의 침윤 및 증가를 억제한 것으로 사료된다. 이를 통해 Ef extract 가 천식 동물모델에서 항염증효과가 있음을 관찰하였지만 Ef extract의 구성 물질 분석 및 작용 메커니즘에 대한 추가연구를 통해 천식 치료물질로의 가능성을 규명할 것이다.

Probiotics are usually defined as intestinal bacteria that provide healthy benefit to the host and may offer new therapeutic materials for the treatment of inflammatory diseases. Lactobacillus, Bifidobacterium and Enterococcus are known as typical probiotics. But, these bacteria have mostly a weak viability and thus decreased probiotics-mediated effects in the intestinal tract. Asthma is an inflammatory airway disease, which is characterized by the releases of inflammatory mediators including cytokine and IgE. They are mainly associated with the recruitment, activation and disregulation of specific inflammatory cells, especially mast cells, monocytes, T cells, eosinophils and neutrophils in asthma. We performed these studies as in vitro and in vivo test the human inflammatory cell lines and ovalbumin (OVA)-induced asthma mouse model. And then the inhibitory effects of Enterococcus faecalis whole extract on inflammatory responses were examined. For our examinations, the E. faecalis whole extract (Ef extract) was acquired from whole bacteria of E. faecalis using freeze/thawing after ultrasonication method. As results, OVA-mediated THP-1 cell viability was decreased by the treatment of Ef extract. In the asthmatic mouse model, Ef extract inhibited the infiltration of inflammatory cells into the inflammatory sites and blood. This whole extract may have anti-asthmatic effects associated with the regulation of IL-5 and IgE expression. It may also be a promising candidate in anti-allergic medicine for the treatment of asthma.

키워드

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