구절초(Chrysanthemum zawadskii var. latilobum)에서 분리된 다당류의 대식세포 면역조절 활성

Abstract

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX> 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-<TEX>${\kappa}B$</TEX>의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다. Macrophages play a pivotal role in the innate and adaptive immune systems. This study investigated the immuno-modulatory activities of polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) in macrophages. Polysaccharides from Chrysanthemum zawadskii var. latilobum were extracted by the ethanol precipitation method. RAW 264.7 mouse macrophage cell line was treated with CZPS (4 to <TEX>$128{\mu}g/mL$</TEX>), and there was no cytotoxicity at a dose below <TEX>$32{\mu}g/mL$</TEX>. The levels of nitric oxide (NO) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-<TEX>${\alpha}$</TEX> and interleukin (IL)-6, IL-<TEX>$1{\beta}$</TEX>) production in the CZPS treated group (<TEX>$32{\mu}g/mL$</TEX>) were <TEX>$6.5{\pm}0.12{\mu}m$</TEX> (NO), <TEX>$1252.8{\pm}79.85$</TEX> (TNF-<TEX>${\alpha}$</TEX>), <TEX>$305.4{\pm}29.41$</TEX> (IL-6), and <TEX>$683.3{\pm}59.71$</TEX> (IL-<TEX>$1{\beta}$</TEX>), respectively, and they were significantly increased when compared to the control group; <TEX>$2.2{\pm}0.03{\mu}m$</TEX> (NO), <TEX>$452.3{\pm}38.34$</TEX> (TNF-<TEX>${\alpha}$</TEX>), <TEX>$31.7{\pm}5.75$</TEX> (IL-6), and <TEX>$184.1{\pm}11.52$</TEX> (IL-<TEX>$1{\beta}$</TEX>). Additionally, protein expression of inducible nitric oxide synthase (iNOS) and phosphorylation of MAPKs and NF-<TEX>${\kappa}B$</TEX> expression were significantly increased upon CZPS treatment. Therefore, these results indicated that polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) may have a potential immunomodulatory activity in macrophages through MAPKs and NF-<TEX>${\kappa}B$</TEX> signaling, and this information is useful for the development of immune enhancing adjuvant materials using a natural ingredient.