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Anti-inflammatory and immune enhancing activities of PB203 in mouse macrophage RAW264.7 cells KCI 등재

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충북대학교 동물의학연구소 (Research Institute of Veterinary Medicine, Chungbuk National University)
초록

Macrophages secrete various cytokines and inflammatory mediators, resulting in playing critical roles in inflammation and immunity. In this study, we investigated anti-inflammatory and immune enhancing properties of PB203, which is a water-soluble extract powder from the fruit of Actinidia polygama, in macrophages. A. polygama is a medicinal plant traditionally known to treat abdominal pain, stroke and rheumatoid arthritis. However, the molecular mechanism for the immune modulation of PB203 is still unclear. Therefore, we assessed the effects of PB203 on the lipopolysaccharide (LPS)-induced inflammation and immune activation, and elucidated its action mechanism in mouse macrophage, RAW264.7 cells. PB203 significantly suppressed not only the levels of nitric oxide (NO), prostaglandin E2, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), but also the mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-α and IL-1β in LPS-stimulated RAW264.7 cells. We also found that these anti-inflammatory activities of PB203 were mediated through the inhibition of toll-like receptor 4 and nuclear factor kappa B (NF-κB) induced by LPS. On the other hand, in normal macrophages, PB203 dose-dependently elevated the gene expression of immunomodulators including granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1 and TNF-α in a statistically significant manner. The expression of IL-10, IL-1β, IL-6, and interferon-γ were also remarkably upregulated by the treatment of 500 μg/mL PB203. In addition, PB203-mediated production of NO and TNF-α was attenuated by NF-κB inhibition in RAW264.7 cells. Interestingly, PB203 promoted the production of nuclear factor erythroid-2-related factor 2, resulting in the increased level of heme oxygenase-1, which is a representative antioxidant enzyme, in both LPS-stimulated and normal RAW264.7 cells. Taken all together, these results suggest that PB203 may have great potential as the candidate of anti-inflammatory agent for improving inflammatory diseases or immune enhancing agent for preventing infectious diseases. Keywords: Actinidia polygama extract (PB203); macrophages; immunomodulator; nuclear factor kappa B (NF-κB); heme oxygenase-1 (HO-1)

목차
INTRODUCTION
MATERIALS AND METHODS
    추출물 준비
    RAW264.7 세포의 배양
    세포독성 측정
    항염증 효과 측정
    면역증강 효과 측정
    qRT-PCR(quantitative real-time polymerase chain reaction)
    NO(nitric oxide), PGE2(prostaglandin E2), TNF-α(tumor necrosis factor-α), IL-1β(interleukin-1β), IL-6(interleukin-6) 생성 측정
    통계 분석
RESULTS
    PB203이 LPS(lipopolysaccharide)로 자극한 대식세포의 생존율 및 항산화 효소 발현에 미치는 효과
    PB203이 대식세포의 염증 매개 인자(inflammatory mediator) 생성에 미치는 영향
    PB203이 대식세포의 전염증성 사이토카인(proinflammatory cytokine) 생성에 미치는 영향
    PB203이 LPS(lipopolysaccharide) 자극에 대한 TLR4(toll-like receptor 4)/NF-κB(nuclearfactor kappa B) 신호전달경로에 미치는 효과
    PB203이 정상 대식세포의 생존율 및 항산화 효소 발현에 미치는 효과
    PB203이 정상 대식세포의 선천 면역 관련 사이토카인 생성에 미치는 영향
    PB203이 정상 대식세포의 TLR4(toll-like receptor 4)/NF-κB(nuclear factor kappa B) 신호전달경로에 미치는 효과
DISCUSSION
저자
  • Seong-Hyun Ho(R&D Center, G&P Bioscience Co., Ltd., Seoul 07528, Korea)
  • Dayoung Kim(R&D Center, G&P Bioscience Co., Ltd., Seoul 07528, Korea)
  • Su-Jin Park(R&D Center, G&P Bioscience Co., Ltd., Seoul 07528, Korea) Corresponding author