Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Anti-inflammatory Activities of Ethanol Extracts from Leaf, Seed, and Seedpod of Nelumbo nucifera

연잎, 연자육, 연자방 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 : 2018.07.31
  • Accepted : 2018.10.26
  • Published : 2019.04.30
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Abstract

Nelumbo nucifera, also known as sacred lotus, has mainly been used as a food throughout the Asian countries. In the present study, we prepared ethanol extracts from leaf (NL), seed (NS), and seedpod (NSP) of Nelumbo nucifera and investigated their anti-inflammatory activities in mouse macrophage RAW 264.7 cells. To evaluate the anti-inflammatory activities of NL, NS, and NSP, nitric oxide (NO) production was measured in LPS-stimulated RAW 264.7 cells. NL, NS, and NSP significantly reduced NO production in a dose-dependent manner without affecting cell viabilities. NL, NS, and NSP dramatically decreased the protein expression of pro-inflammatory genes such as iNOS and COX-2. NL, NS, and NSP also suppressed phosphorylation of MAPKs and the nuclear translocation of $NF-{\kappa}B$ p65 indicating they have their anti-inflammatory activities via regulating mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B ($NF-{\kappa}B$) pathways. In addition, we analyzed the production of reactive oxygen species (ROS) by the treatment of NL, NS, and NSP. All extracts reduced ROS production in a dose-dependent manner. And also, they increased heme oxygenase-1 (HO-1) protein expression and the nuclear translocation of nuclear respiratory factor 2 (Nrf2). In conclusion, our results suggest that Nelumbo nucifera has its anti-inflammatory activity via regulating MAPKs, $NF-{\kappa}B$, and Nrf2/HO-1 pathways.

연은 아시아 국가에서 음식과 약재로 널리 사용되는 재료이다. 본 연구에서는 연의 잎(leaf, NL), 자육(seed, NS), 자방(seedpod, NSP)으로부터 에탄올 추출물을 제조하고 이들의 항염증 활성과 작용기전을 규명하였다. 이들의 항염증 활성을 연구하기 위하여 LPS로 자극된 RAW 264.7 세포에서 nitric oxide (NO) 생산을 측정하였다. NL, NS, NSP는 세포 생존율에 영향을 주지 않으면서, 농도의존적으로 NO의 생산을 현저하게 저해하였으며, iNOS 및 COX-2와 같은 pro-inflammatory 중재자들의 단백질 발현을 감소시켰다. 또한, NL, NS, NSP는 MAPKs 단백질의 인신화를 감소시키고 $NF-{\kappa}B$ p65의 핵으로의 이동을 저해함으로써, 세 추출물에 의한 항염증 활성은 MAPKs 경로와 $NF-{\kappa}B$ 경로를 조절함으로써 이루어짐을 제시한다. 게다가, ROS의 생성이 세 추출물에 의해서 모두 저해되었으며, HO-1의 발현과 HO-1의 전사조절인자인 Nrf2의 핵으로의 이동이 증가되었다. 결론적으로, 이러한 연구 결과는 연의 다양한 부위의 추출물인 NL, NS 그리고 NSP는 항염증 활성을 가지고 있으며, MAPKs, $NF-{\kappa}B$, Nrf2/HO-1 등 다양한 신호경로를 통해 조절할 수 있음을 제시한다.

Keywords

SMGHBM_2019_v29n4_436_f0001.png 이미지

Fig. 1. Effects of leaf (NL), seed (NS) and seedpod (NSP) extracts of Nelumbo nucifera on nitric oxide (NO) production and cell viability in LPS-stimulated RAW264.7 macrophage.

SMGHBM_2019_v29n4_436_f0002.png 이미지

Fig. 2. Down-regulation of iNOS and COX-2 proteins by the treatment of NL, NS and NSP.

SMGHBM_2019_v29n4_436_f0003.png 이미지

Fig. 3. Inhibition of phosphorylation of MAPKs and nuclear translocation of NF-κB p65 by the treatment of NL, NS and NSP.

SMGHBM_2019_v29n4_436_f0004.png 이미지

Fig. 4. ROS scavenging effects of NL, NS, and NSP extracts.

SMGHBM_2019_v29n4_436_f0005.png 이미지

Fig. 5. Up-regulation of HO-1 via increased nuclear translocation of Nrf2 by NL, NS, and NSP treatment.

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