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RAW 264.7세포에서 lipopolysaccharide로 유발시킨 염증반응에 대한 Bulnesia sarmienti 열수추출물의 억제효과

Bulnesia Sarmienti Aqueous Extract Inhibits Inflammation in LPS-Stimulated RAW 264.7 Cells

  • 전용필 (성신여자대학교 자연과학대학) ;
  • ;
  • 박창호 ((재)대구테크노파크 바이오산업지원센터) ;
  • 홍주헌 ((재)대구테크노파크 바이오산업지원센터) ;
  • 이기동 ((재)대구테크노파크 바이오산업지원센터) ;
  • 송재찬 (경북대학교 수의과대학) ;
  • 김길수 (경북대학교 수의과대학)
  • 발행 : 2009.04.30

초록

본 연구는 BS 열수추출물의 항염증 활성을 조사하기 위하여 LPS로 자극된 RAW 264.7세포에서 세포가 방출하는 NO, $PGE_2$, COX생성량과 염증성 사이토카인인 $IL-1{\beta}$, IL-6, $TNF-{\alpha}$의 생성량의 변화를 조사하여 항염증성 기능의 유무와 정도를 알아보고자 하였다. 마우스 대식세포인 RAW 264.7에 대한 BS추출물의 농도별(0, 50, 100, $200\;{\mu}g/ml$) 세포독성을 알아보기 위하여 MTT assay를 수행하여 $200\;{\mu}g/ml$ 이하의 농도에서는 세포독성이 낮아 기본적으로 세포의 생존율에 영향을 주지 않는다는 사실을 알 수 있었다. RAW 264.7세포에 BS추출물을 전처리하고 LPS를 18시간 동안 처리한 후 NO의 생성량은 정상세포에 비하여 약 8배 이상 증가되었으며, BS추출물을 $100\;{\mu}g/ml$ 농도 이상으로 처리한 실험군에서는 농도의존적으로 nitrite의 생성량이 유의하게 감소되었다. $PGE_2$의 생성량은 LPS처리에 의하여 정상세포에 비하여 약 14배 이상 증가되었으며 BS추출물을 100 및 $200\;{\mu}g/ml$의 농도로 처리한 실험군에서는 농도의존적으로 유의하게 각각 55.1%, 39.9% 감소되었다. COX의 생성량은 LPS처리에 의하여 정상세포에 비하여 약 5.6배 증가되었으며 $100\;{\mu}g/ml$ 이상의 농도처리군에서는 농도의존적으로 유의하게 감소되었다. LPS처리후 $IL-1{\beta}$, IL-6, $NF-{\alpha}$의 생성량은 정상세포에 비하여 급격하게 증가되었으며, 여러 농도(50, 100, $200\;{\mu}g/ml$)의 BS추출물로 처리한 실험군에서는 각각 아무런 처치를 하지 않은 세포에 비하여 농도의존적인 각각의 생성량이 감소되었다. 특히 $100\;{\mu}g/ml$ 농도 이상의 BS처리군에서는 모두 통계학적 유의성이 인정되었다. 이러한 결과는 BS에서 염증억제 효과가 있음을 in vitro 연구에서 확인할 수 있었고, 향후 유효성분 추출을 통한 항염증 물질의 연구 또는 예방하거나 치료할 수 있는 염증 억제 성분의 분리 및 그 작용기전 연구에 중요한 기초 자료가 될 것이라 사료된다.

Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. We investigated the cytotoxic activities and the inhibitory effects of BS bark extract(0, 50, 100 and $200\;{\mu}g/\;mL$) on the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase (COX) and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) in the lipopolysaccharide (LPS) (100 ng/ml)-stimulated murine macrophage cell line RAW264.7. The levels of NO, COX, PGE2 production and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) were measured by ELISA kit. Cell viability, as measured by the MTT assay, showed that BS extract had no significant cytotoxicity in RAW264.7 cells. BS extract significantly inhibited the LPS-induced NO, $PGE_2$ and COX production accompanied by an attenuation of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ formation in macrophages. These results suggest that BS extract has potential as an herbal medicine for the treatment of inflammatory diseases.

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참고문헌

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