This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

Abstract
서 론
재료 및 방법
    1. 시료의 추출
    2. 구성당 분석
    3. RAW 264.7 cell에 대한 NO 및 cytokine 생성능
    4. qPCR을 통한 유전자 발현도 측정
    5. Western blotting
    6. Confocal을 이용한 NF-κB 경로의 확인
    7. 대식세포의 pattern recognition receptors (PRRs)
    8. 통계처리
결과 및 고찰
    1. 진피로부터 분리한 다당의 구성당 분석
    2. 진피 유래 조다당의 NO 및 cytokine 생성능
    3. 진피 유래 조다당의 mRNA gene expression
    4. 진피 유래 조다당의 중화항체를 이용한 대식세포의 수용체 경로 확인
    5. 진피 유래 조다당의 대식세포 내 MAPK 및 NF-κB 경로의 활성화
요약 및 결론
References

• 이경애(안양대학교 식품영양학과) | Kyung-Ae Lee (Dept. of Food and Nutrition, Anyang University)
• 박혜령(경희대학교 한방생명공학과) | Hye-Ryung Park (Graduate School of Biotechnology, and College of Life Science, Kyung Hee University) Corresponding author