Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Anti-inflammatory Activities of Ethanol Extracts of Dried Lettuce (Lactuca sativa L.)

건조 상추 에탄올 추출물의 항염증 활성

  • Lee, Eun-Joo (Department of Biological Sciences, Andong National University) ;
  • Seo, Yu-Mi (Department of Biological Sciences, Andong National University) ;
  • Kim, Yong-Hyun (Department of Biological Sciences, Andong National University) ;
  • Chung, Chungwook (Department of Biological Sciences, Andong National University) ;
  • Sung, Hwa-Jung (Department of Food and Nutrition, Andong National University) ;
  • Sohn, Ho-Yong (Department of Food and Nutrition, Andong National University) ;
  • Park, Jong-Yi (Gyeongbuk Institute for Bio Industry) ;
  • Kim, Jong-Sik (Department of Biological Sciences, Andong National University)
  • 이은주 (국립안동대학교 생명과학과) ;
  • 서유미 (국립안동대학교 생명과학과) ;
  • 김용현 (국립안동대학교 생명과학과) ;
  • 정정욱 (국립안동대학교 생명과학과) ;
  • 성화정 (국립안동대학교 식품영양학과) ;
  • 손호용 (국립안동대학교 식품영양학과) ;
  • 박종이 (경북바이오산업 연구원) ;
  • 김종식 (국립안동대학교 생명과학과)
  • Received : 2018.12.19
  • Accepted : 2019.01.28
  • Published : 2019.03.30
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Abstract

Lettuce (Lactuca sativa L.) is one of the most popular green leafy vegetables, and it contains various beneficial components including polyphenolic compounds and has been known to possess various biological functions such as anti-microbial, anti-oxidative, and anti-inflammatory activities. In the present study, we prepared ethanol extract of dried lettuce (DLE) and investigated its anti-inflammatory activity. To evaluate the anti-inflammatory activity of DLE, nitric oxide (NO) production was measured in LPS-activated mouse macrophage RAW 264.7 cells. DLE significantly suppressed NO production in these cells without affecting cell viabilities while resveratrol was used as a positive control. DLE dramatically decreased the expression of pro-inflammatory genes such as iNOS and COX-2 at the mRNA and protein levels and reduced the expression of several cytokines including $IL-1{\alpha}$, $IL-1{\beta}$, IL-1F6, $TNF-{\alpha}$, CSF2 and CXCL10. In addition, DLE suppressed phosphorylation of MAPKs and the nuclear translocation of $NF-{\kappa}B$ p65 indicating DLE shows its anti-inflammatory activity via regulating MAPKs pathway and $NF-{\kappa}B$ pathways. And also, DLE reduced the production of reactive oxygen species in a dose-dependent manner. DLE increased HO-1 protein expression, and also increased the nuclear translocation of Nrf2. Overall, our results suggest that lettuce down-regulate various pro-inflammatory genes and have its anti-inflammatory activity via regulating MAPKs, $NF-{\kappa}B$, and Nrf2/HO-1 pathways.

상추는 가장 선호하는 녹색 채소 중 하나이다. 상추는 폴리페놀성 화합물을 비롯한 다양한 성분을 함유하고 있으며, 항균, 항산화, 항염증 등의 생리활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 건조상추의 에탄올 추출물(DLE)을 제조하고 이들의 항염증 활성을 연구하였다. DLE의 항염증 활성을 측정하기 위하여 LPS로 활성화된 마우스 대식세포 RAW 264.7 세포주에서 nitric oxide (NO) 생성을 측정하였다. DLE는 세포주의 생존에는 영향을 미치지 않으면서 NO 생산을 현저하게 저해하였다. DLE에 의해 염증 유전자인 iNOS와 COX-2의 유전자와 단백질의 발현이 모두 감소하였으며, 6개의 염증관련 cytokine 유전자($IL-1{\alpha}$, $IL-1{\beta}$, IL-1F6, $TNF-{\alpha}$, CSF2, 그리고 CXCL10)의 발현이 모두 감소하였다. 또한, DLE의 처리는 MAPKs 경로의 인산화를 모두 저해하였으며, $NF-{\kappa}B$ p65의 핵으로의 이동을 저해하였다. 이러한 결과는 DLE의 항염증 활성은 MAPKs 경로와 $NF-{\kappa}B$ 경로를 조절함으로써 이루어짐을 시사한다. 또한, DLE는 농도의존적으로 reactive oxygen species (ROS)의 생산을 저해하였으며, hemeoxygenase-1 (HO-1) 단백질의 발현을 증가시켰으며, HO-1의 전사조절인자인 Nrf2의 핵으로의 이동을 증가시켰다. 결론적으로, 이러한 연구결과는 DLE가 염증관련 유전자의 발현을 감소시키며, MAPKs, $NF-{\kappa}B$, 그리고 Nrf2/HO-1 등 다양한 경로를 조절함으로써 항염증 활성을 가지는 것을 제시한다.

Keywords

SMGHBM_2019_v29n3_325_f0001.png 이미지

Fig. 1. Effects of Dry-Lettuce ethanol extract (DLE) on nitric oxide (NO) production and cell viability in LPS-stimulated RAW 264.7 macrophage.

SMGHBM_2019_v29n3_325_f0002.png 이미지

Fig. 2. Down-regulation of pro-inflammatory mediators such as iNOS and COX-2 genes by the treatment of DLE.

SMGHBM_2019_v29n3_325_f0003.png 이미지

Fig. 3. Inhibition of phosphorylation of MAPKs and the nuclear translocation of NF-κB by DLE treatment.

SMGHBM_2019_v29n3_325_f0004.png 이미지

Fig. 4. ROS scavenging effects of DLE and up-regulation of HO-1 via increased nuclear translocation of Nrf2 by DLE treatment.

Table 1. Sequences of oligonucleotide primers used for reverse-transcription PCR

SMGHBM_2019_v29n3_325_t0001.png 이미지

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