화학공학소재연구정보센터
홈 로그인 로그아웃 연락처 사이트맵
Applied Chemistry for Engineering, Vol.30, No.2, 145-150, April, 2019
DOI10.14478/ace.2018.1102  Export Citation
추출 용매에 따른 Lactobacillus plantarum 발효 우슬의 항염증 효과 증진
Enhancement of Anti-inflammatory Activity of Lactobacillus plantarum Fermented by Achyranthes japonica on Extraction Solvents
조은솔1, 우영민1, 김옥주1, 조민영2, 안미영3, 이재화2, 3, 하종명2, 3, 김안드레2, 3, †
  • 1신라대학교 자연과학연구소
  • 2신라대학교 일반대학원 바이오과학과
  • 3신라대학교 제약공학과
Eun Sol Jo1, Young Min Woo1, Ok Ju Kim1, Min Young Jo2, Mee Young Ahn3, Jae-Hwa Lee2, 3, Jong-Myung Ha2, 3, and Andre Kim2, 3, †
  • 1Natural Science Institute, Silla University, Busan 46958, Korea
  • 2Department of Bioscience, Graduate School of Silla University, Busan 46958, Korea
  • 3Department of Pharmaceutical Engineering, Silla University, Busan 46958, Korea
E-mail:
초록
본 연구에서는 우슬(Achyranthes japonica, AJ)과 Lactobacillus plantarum으로 발효한 우슬(AJ-LP)을 5가지 용매(물, 에탄올, 헥산, 아세트산에틸, 부탄올)로 추출하여 RAW264.7 세포에서 항염증 활성에 미치는 영향을 확인하였다. lipopolysaccharide (LPS)로 유도된 RAW264.7 세포에서 nitric oxide (NO) 및 cytokine 생성을 측정하였으며 western blot으로 cyclooxygenase-2 (COX-2)와 nitric oxide synthase (iNOS)의 발현 정도를 관찰하였다. 세포 독성은 CCK assay를 통해 확인하였으며 독성을 가지지 않는 농도인 100 μg/mL로 실험을 진행하였다. NO 생성 저해 활성 결과 AJ-LP를 에탄올(E)로 추출한 시료가 LPS만 처리한 control군에 비해 약 74%로 저해율이 가장 높았으며 염증관련 cytokine인 Interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and Interleukin-1β (IL-1β)의 결과에서도 약 57, 70, 74%의 저해율로 우수한 효능을 보였다. COX-2와 iNOS 발현결과 AJ군에서 대조군(20-hydroxyecdysone)의 저해율이 가장 높은 것으로 나타났다. AJ-LP군에서 COX-2 발현량은 헥산(H)으로 추출한 시료가 대조군에 비해 약 16%로 감소하였고 iNOS 발현량은 부탄올(B)으로 추출한 시료가약 7%로 감소하였다. 이상의 결과들에서 우슬보다 L. plantarum으로 발효한 우슬이 천연물 소재로서의 활용 가능성이 높음을 시사하였다.
In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of 100 μg/mL, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and Interleukin-1β (IL-1β), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.
Keywords:RAW264.7;Anti-inflammatory;Nitric oxide;Achyranthes japonica;Lactobacillus plantarum
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