Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Protective effect of Eucommia ulmoides oliver leaves against PM2.5-induced oxidative stress in neuronal cells in vitro

미세먼지(PM2.5)로 유도된 산화적 스트레스에 대한 두충(Eucommia ulmoides Oliver) 잎의 in vitro 뇌 신경세포 보호 효과

  • Kim, Min Ji (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kang, Jin Yong (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Park, Seon Kyeong (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Jong Min (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Moon, Jong Hyun (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Gil Han (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Lee, Hyo Lim (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Jeong, Hye Rin (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Heo, Ho Jin (Division of Applied Life Science (BK21), Institute of Agriculture and Life Science, Gyeongsang National University)
  • 김민지 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 강진용 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 박선경 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 김종민 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 문종현 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 김길한 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 이효림 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 정혜린 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원) ;
  • 허호진 (경상국립대학교 응용생명과학부(BK21), 농업생명과학연구원)
  • Received : 2021.04.30
  • Accepted : 2021.06.16
  • Published : 2021.08.31
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Abstract

This study was performed to examine the neuroprotective effect of the ethyl acetate fraction from Eucommia ulmoides oliver leaf (EFEL) on PM2.5-induced cytotoxicity. EFEL had higher total phenolic and flavonoid contents than the other fractions. In ABTS and DPPH radical scavenging activities, the IC50 of EFEL was measured as 212.80 and 359.13 ㎍/mL, respectively. To investigate the neuroprotective effect of EFEL, MTT and DCF-DA assays were performed on HT22, MC-IXC, and BV-2 cells. EFEL effectively decreased PM2.5-induced intercellular reactive oxygen species (ROS) content and inhibited PM2.5-induced cell death. In the results of protein expression related to cellular cytotoxicity on microglial cells (BV-2), EFEL had an improvement effect on cell apoptosis and inflammatory pathways. Rutin and chlorogenic acid were identified as the main physiological compounds. Moreover, it was expected that EFEL, including rutin and chlorogenic acid, could be functional food substances with neuroprotective effects against PM2.5-induced oxidative stress.

본 연구에서는 두충 잎의 분획물을 이용하여 미세먼지(PM2.5)로 유도한 in vitro 뇌 신경세포 독성에 대한 항산화 활성 및 세포보호 효과를 확인하였다. EFEL은 우수한 ABTS 및 DPPH 라디칼 소거능을 나타냈으며, 지질과산화물 억제활성에서 양성대조군인 catechin과 동일한 농도에서 유사한 활성을 나타냈다. 또한, EFEL은 미세먼지로 유도된 해마세포(HT22), 뇌신경세포(MC-IXC) 및 미세아교세포(BV-2) 내의 ROS 수준을 효과적으로 감소시켜주었으며, 세포 생존능을 증가시킴으로써 세포보호 효과를 나타냈다. 미세먼지로 활성화된 BV-2에서 EFEL은 세포 활성화와 관련된 세포막 수용체인 TLR4와 p-JNK의 발현을 감소시켰다. 또한, 세포 사멸에 관여하는 caspase-3 활성화 전 단계인 procaspase-3와 세포 생존 경로에 관여하는 p-Akt 발현의 증가된 수준을 나타냈며, 염증성 사이토카인으로써 증가된 TNF-α의 수준을 감소시켰다. EFEL의 생리활성 물질을 HPLC로 분석한 결과, rutin과 chlorogenic acid가 확인되었다. 결과적으로, 두충 잎의 아세트산에틸 분획물은 우수한 페놀성 화합물 및 플라보노이드 함량을 나타냄으로써 높은 항산화능을 보였으며, 미세먼지로 유도된 산화적 스트레스에서 사멸 및 염증반응을 조절하여 뇌 신경세포를 보호하였다. 이를 고려할 때, EFEL은 미세먼지로 유도된 염증성 뇌신경세포 관련 질환 예방에 도움을 줄 수 있는 건강기능식품 소재로 활용될 수 있을 것으로 판단된다.

Keywords

Acknowledgement

본 연구는 한국연구재단의 개인기초연구(2018R1D1A3B07043398)의 지원을 받아 수행된 결과로 이에 감사드립니다.

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