[논문]망개나무 추출물의 NF-κB 및 MAPK 신호전달 억제를 통한 항염증 효과

김다솜; 어현지; 강연경; 박광훈

doi:10.7732/kjpr.2021.34.1.031

망개나무 추출물의 NF-κB 및 MAPK 신호전달 억제를 통한 항염증 효과
Anti-inflammatory Effect of Berchemia berchemiaefolia Leaves through Inhibition of NF-κB and MAPK Signaling Activation in LPS-stimulated RAW264.7 Cells 원문보기

韓國資源植物學會誌 = Korean journal of plant resources, v.34 no.1, 2021년, pp.31 - 36

김다솜 (국립산림과학원 산림약용자원연구소) , 어현지 (국립산림과학원 산림약용자원연구소) , 강연경 (국립산림과학원 산림약용자원연구소) , 박광훈 (국립산림과학원 산림약용자원연구소)

초록
AI-Helper

본 연구는 망개나무 추출물의 항염증 효과를 구명하기 위하여 수행되었다. 망개나무를 이용한 다양한 기능성 연구가 진행되고 있으나 망개나무 항염증에 대한 연구는 과학적 근거가 부족한 실정이다. 따라서 망개나무 잎(BBK-L) 추출물이 마우스 대식세포인 RAW264.7 세포에서 LPS에 의한 염증반응에 미치는 영향을 평가하였다. LPS로 유도된 마우스 대식세포 RAW264.7에 BBK-L 추출물을 농도별로 처리하였을 때, NO 생성량과 만성염증 유발인자인 iNOS, COX-2, IL-1β, TNF-α및 IL-6 발현 감소를 확인하였으며, NF-κB 및 MAPK의 인산화를 억제하는 것을 확인하였다. 이러한 연구결과를 토대로 BBK-L 추출물이 만성 염증질환의 예방과 치료 또는 기능성 소재로 활용될 수 있는 기초적인 정보를 제공할 수 있을 것으로 사료된다.

Abstract ▼ AI-Helper

Berchemia berchemiaefolia (Makino) Koidz. is found not only in korea, but also in japan and is known as a rare plant all over the world. In this study, we evaluated anti-inflammatory effect of B. berchemiaefolia (Makino) Koidz. leaves (BBK-L) in LPS-stimulated RAW264.7 cells. BBK-L inhibited the generation of NO through the suppression of iNOS experession. BBK-L attenuated the expression of iNOS, COX-2, TNF-α, IL-1β, IL-6 induced by LPS. BBK-L decreased LPS-mediated IκB-α degradation inhibite which resulted in the inhibition of NF-κB activation in RAW264.7 cells. In addition, BBK-L suppressed ERK1/2, JNK phosphorylation induced by LPS. BBK-L showed anti-inflammatory effect through blocking the generation of the inflammatory mediators such as NO, iNOS, COX-2, TNF-α, IL-1β, IL-6 via the inhibiting of NF-κB and MAPK signaling activation. These results suggests that BBK-L may have great potential for the development of anti-inflammatory drug to treat acute and chronic inflammatory disorders.

주제어

표/그림 (5)

표 Table 1. Sequence of oligonucleotide primers used for RT-PCR
그림 Fig. 1. Inhibitory effect of the extracts from leaves (BBK-L), branches (BBK-B) and fruits (BBK-F) of BBK against NO production in LPS-stimulated RAW264.7 cells. (A) RAW264.7 cells were pretreated with BBK-L, BBK-B and BBK-F for 6 hours and then co-treated with LPS (1 ㎍/mL) for 18 hours. (B) RAW264.7 cells were pretreated with BBK-L (0, 12.5, 25 and 50 ㎍/mL) for 6 hours and then co-treated with LPS (1 ㎍/mL) for 18 hours. NO production was measured by Griess assay. (C) RAW264.7 cells were treated with BBK-L for 24 h. Cell viability was measured using MTT assay. #p < 0.05 compared to the cells without the treatment, and *p < 0.05 compared to the cells treated with LPS alone.
그림 Fig. 2. Effect of BBK-L on the expression of iNOS, COX-2, TNF-α, IL-1β and IL-6 in LPS-stimulated RAW264.7 cells. RAW264.7 cells were pre-treated with BBK-L for 6 hours and then co-treated with LPS (1 μg/mL) for 18 hours. For RT-PCR analysis, total RNA was prepared. GAPDH was used as internal control for RT-PCR. #p < 0.05 compared to the cells without the treatment, and *p < 0.05 compared to the cells treated with LPS alone.
그림 Fig. 3. Effects of BBK-L on NF-κB signaling activation in LPS-stimulated RAW264.7 cells (A) RAW264.7 cells were pretreated with BBK-L (0, 12.5, 25, 50 μg/mL) for 6 hours and then co-treated with LPS (1 μg/mL) for 40 minutes. (A and B) RAW264.7 cells were pretreated for 6 hours with the indicated concentrations of BBK-L (12.5, 25, 50 μg/mL) and than co-treated LPS (1 μg/mL) treatment for 40 minutes. (C) RAW264.7 cells were pre-treated with BBK-L (0, 12.5, 25, 50 μg/mL) for 40 minutes. After the treatment, the cytosol and nucleus were prepared. For Western blot analysis, the cell lysates were subjected to SDS-PAGE and the Western blot was performed using antibodies against IκB-α and p-IκB-α and nucleus p65. β-actin was used as a internal control. #p < 0.05 compared to the cells without the treatment, and *p < 0.05 compared to the cells treated with LPS alone.
그림 Fig. 4. Effects of BBK-L on MAPK signaling activation in LPS-stimulated RAW264.7 cells. RAW264.7 cells were pretreated with BBK-L (0, 12.5, 25, 50 μg/mL) for 6 hours and then co-treated with LPS (1 μg/mL) for 40 minutes. For western blot analysis, the cell lysates were subjected to SDS-PAGE and the western blot was performed using antibodies against p-p38, total-ERK1/2, p-ERK1/2, total p-38, p-JNK and total-JNK. #p < 0.05 compared to the cells without the treatment, and *p < 0.05 compared to the cells treated with LPS alone.

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