본 연구는 편백나무 잎 분획물의 항염증 기능성 소재로서의 활용 가능성을 평가하기 위해 99% 에탄올로 추출한 편백잎 추출물 (CO99EL)을 헥산 (CO99EL-H), 클로로포름 (CO99EL-C), 에틸 아세테이트 (CO99EL-E), 부탄올 (CO99EL-B)과 증류수 (CO99EL-W) 순서대로 분획하였다. 각각의 분획물의 항염증 효과는 LPS로 유도된 RAW264.7 마우스 대식세포를 이용하여 수행하였다. 세포독성은 CO99EL-H와 CO99EL-C에서 가장 높았으며 CO99EL-W에서 가장 낮음을 확인하였다. 흥미롭게도, LPS로 유도된 iNOS의 발현과 NO의 생산은 CO99EL-H와 CO99EL-E에 의해 현저하게 감소하였고, COX-2의 발현은 CO99EL-B와 CO99EL-W에 의해 현저하게 감소하였다. 또한, LPS에 의해 증가된 염증성 사이토카인인 interleukin(IL)-1β는 CO99EL-C, CO99EL-E, CO99EL-B와 CO99EL-W에 의 해 현저하게 감소하였고, IL-6는 CO99EL-B와 CO99EL-W에 의해 현저하게 감소하였다. 그뿐만 아니 라, LPS에 의해 활성화된 janus kinase (JAK)/signaling transducer and activator of transcription (STAT) 신호 전달 경로는 CO99EL-H와 CO99EL-C에 의해 상당히 감소하였고, mitogen-activated protein kinase (MAPK)은 CO99EL-C에 의해 약간 감소하였다. 하지만, nuclear factor (NF)-κB의 활 성은 어떤 분획물도 감소시키지 못했다. 본 연구의 결과를 통해, CO99EL의 분획물들은 분획에 사용되 는 용매에 따라 항염증 작용기전이 다름을 확인하였다.

In this study, to evaluate the possibility of utilizing Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) leaf fractions as anti-inflammatory functional materials, C. obtusa extract extracted with 99% ethanol (CO99EL) was fractionated with hexane (CO99EL-H), chloroform (CO99EL-C), ethyl acetate (CO99EL-E), butanol (CO99EL-B) and distilled water (CO99EL-W). The anti-inflammatory effects of each fraction was performed using lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophages. Cytotoxicity was highest in CO99EL-H and CO99EL-C and lowest in CO99EL-W. Interestingly, LPS-induced iNOS expression and NO production were significantly reduced by CO99EL-H and CO99EL-E, and COX-2 expression was significantly reduced by CO99EL-B and CO99EL-W. In addition, interleukin (IL)-1β, an inflammatory cytokine increased by LPS, was significantly reduced by CO99EL-C, CO99EL-E, CO99EL-B and CO99EL-W, and IL-6 was significantly reduced by CO99EL-B and CO99EL-W. Therefore, the janus kinase (JAK)/signaling transducer and activator of transcription (STAT) signaling pathway activated by LPS was significantly reduced by CO99EL-H and CO99EL-C, and the mitogen-activated protein kinase (MAPK) signaling pathway was slightly reduced by CO99EL-H and CO99EL-C. However, nuclear factor (NF)-κB activity was not reduced by any fractions. Based on the results of this study, it was confirmed that CO99EL fractions have different anti-inflammatory mechanisms depending on the solvent used for fractionation.

1. 서 론
2. 실 험
    2.1. 실험재료
    2.2. 항체
    2.3. 편백나무 추출 및 분획
    2.4. 세포배양
    2.5. 세포독성 분석
    2.6. NO 생산 측정
    2.7. Western blotting
    2.7. 실시간 중합효소연쇄반응 (qPCR)
    2.8. 통계분석
3. 결과 및 고찰
    3.1. 편백 분획물 분획과정 및 세포독성 평가
    3.2. LPS에 의해 유도된 iNOS 및 COX-2발현에 대한 분획물의 효과
    3.3. LPS에 의해 유도된 JAK/STAT 활성화와IL 발현에 대한 분획물의 효과
    3.4. LPS에 의해 유도된 JAK/STAT 활성화와IL 발현에 대한 분획물의 효과
4. 결 론
References

• 권용진(경성대학교 화장품학과) | Yong-Jin Kwon (Department of Cosmetic Science, Kyungsung University, Busan, 48434, South Korea) Corresponding author