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RAW264.7 세포주를 통한 복분자 30% 에탄올 추출물의 면역기능 활성증진효과 검토

Immune enhancement effect of Rubus coreanus Miquel (RC) (30% EtOH extract) in RAW264.7 cells

고성규|신용철|조재경|최한석|김용국|김치홍|김민수
대한예방한의학회 (Society of Preventive Korean Medicine)|2012.12|대한예방한의학회지 (Society of Preventive Korean Medicine)|제16권 제3호|pp. 155-165

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참고문헌

키워드

Rubus coreanus Miquel (RC)nitric oxide (NO)reactive oxygen species (ROS)cyclooxygenase-2 (COX-2)inductible nitric oxide (iNOS)mitogen-activated protein kinase (MAPK)
주제분류한의학

초록

Objective : The object of this study is to confirm the immune enhancement effect of Rubus coreanus Miquel (RC) (30% EtOH extract) on RAW264.7 mouse macrophage cell line. Methods : RAW264.7 cells were treated with 10-500μg/mL RC for 24 hours. Cell viability was then measured using WST assays. Levels of intracellular NO and ROS were measured by Griess reagent and DCFH-DA staining respectively. Levels of iNOS and COX-2 mRNA was determined by RT-PCR. Secretion levels of IL-1β and IL-6 cytokines were evaluated by sandwich ELISA assay. Western blot analysis was performed to measure the levels of intracellular molecules related to MAPKs pathways. Results : RC suppressed the growth of RAW264.7 cells. RC increased the production of NO and ROS. RC increased the mRNA and protein levels of COX-2 and iNOS. RC augmented the levels of secreted IL-1β and IL-6 cytokines. RC increased MAPKs phosphorylation. Conclusion : In summary, our result shows that RC has inflammatory effect increasing the levels of NO, ROS and secreted cytokines and activating MAPKs. Hence, RC seems to have an immune enhancement effect.

Abstract

Objective : The object of this study is to confirm the immune enhancement effect of Rubus coreanus Miquel (RC) (30% EtOH extract) on RAW264.7 mouse macrophage cell line. Methods : RAW264.7 cells were treated with 10-500μg/mL RC for 24 hours. Cell viability was then measured using WST assays. Levels of intracellular NO and ROS were measured by Griess reagent and DCFH-DA staining respectively. Levels of iNOS and COX-2 mRNA was determined by RT-PCR. Secretion levels of IL-1β and IL-6 cytokines were evaluated by sandwich ELISA assay. Western blot analysis was performed to measure the levels of intracellular molecules related to MAPKs pathways. Results : RC suppressed the growth of RAW264.7 cells. RC increased the production of NO and ROS. RC increased the mRNA and protein levels of COX-2 and iNOS. RC augmented the levels of secreted IL-1β and IL-6 cytokines. RC increased MAPKs phosphorylation. Conclusion : In summary, our result shows that RC has inflammatory effect increasing the levels of NO, ROS and secreted cytokines and activating MAPKs. Hence, RC seems to have an immune enhancement effect.

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