We investigated the anti-inflammatory effects of shiitake mushroom and kelp (SMK) mixture extracts in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 cells. Treatment of RAW 264.7 cells with LPS significantly increased NO (nitric oxide) production, pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and IL-1β), and inflammation-related genes (COX-2 and inducible nitric oxide synthase (iNOS)). In cytotoxicity testing using RAW 264.7 cells, SMK mixture extracts in the range of 1-16 μg/mL did not inhibit cell proliferation. However, SMK mixture extracts significantly inhibited NO production in a dose-dependent manner (p<0.05). SMK treatment significantly decreased TNF-α, IL-6, IFN-γ, and IL-1β levels compared to the LPS group, and similarly, pro-inflammatory cytokine mRNA levels also decreased. SMK mixture extracts reduced the mRNA expression of COX-2 and iNOS in RAW 264.7 cells compared to LPS (p<0.05). The above results show that SMK mixture extracts suppressed the inflammatory response induced by LPS. In particular, the extracts were shown to regulate the inflammatory response by suppressing the expression of inflammatory cytokines and inflammation-related enzymes.

서 론
재료 및 방법
    1. 재료 준비
    2. RAW 264.7 세포 배양
    3. 세포독성분석
    4. Nitric oxide(NO) 생성 농도 측정
    5. 전염증성 사이토카인 분비량 측정
    6. 염증관련 사이토카인 mRNA 발현 측정
    7. 통계처리
결과 및 고찰
    1. 세포 생존율 측정
    2. NO 생성량 분석
    3. 전염증성 사이토카인 분비량 측정
    4. 전염증성 사이토카인 mRNA 발현
요약 및 결론
감사의 글
References

• 김수봉(호서대학교 보건산업연구소 연구원) | Soo Bong Kim (Researcher, Institute of Health Industry, Hoseo University, Seoul 06724, Korea)
• 강순아(호서대학교 벤처대학원 융합공학과 교수, 호서대학교 보건산업연구소 소장) | Soon Ah Kang (Professor, Dept. of Convergence Science and Technology, Graduate School of Venture, Hoseo University, Seoul 06724, Korea, Director, Institute of Health Industry, Hoseo University, Seoul 06724, Korea) Corresponding author