Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Neuroprotective Effects of Cirsium setidens, Pleurospermum kamtschaticumin, and Allium victorials Based on Antioxidant and p38 Phosphorylation Inhibitory Activities in SK-N-SH Neuronal Cells

SK-N-SH 신경세포내 항산화 효과와 p38 인산화 억제에 의한 곤드레, 누룩치 그리고 산마늘의 신경 보호 효과

  • Chung, Mi Ja (Department of Food Science and Nutrition, Gwangju University) ;
  • Park, Yong Il (Department of Biotechnology, The Catholic University of Korea) ;
  • Kwon, Ki Han (Department of Food Science and Nutrition, Gwangju University)
  • 정미자 (광주대학교 식품영양학과) ;
  • 박용일 (가톨릭대학교 생명공학과) ;
  • 권기한 (광주대학교 식품영양학과)
  • Received : 2014.11.25
  • Accepted : 2015.01.19
  • Published : 2015.03.31
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Abstract

Oxidative stress is one of the key mechanisms involved in neuronal damage. Neuroprotective effects and underlying mechanisms of action of several wild vegetables, Cirsium setidens (CS), Pleurospermum kamtschaticumin (PK), and Allium victorials (AV), against oxidative stress induced by hydrogen peroxide in SK-N-SH cells were investigated. CS and AV up to $400{\mu}g/mL$ showed no detectable effects on cell viability of human SK-N-SH neuro-blastoma cells compared with control. Incubation of SK-N-SH cells with hydrogen peroxide resulted in significant induction of cell death and reaction oxygen species (ROS) production, whereas treatment of cells with CS and AV significantly reduced cell death and ROS production, respectively. Among the wild vegetables tested, CS and PK showed more effective DPPH radical scavenging activity than AV, whereas PK showed strong cytotoxicity in SK-N-SH cells compared with the control. CS showed much higher inhibitory effects on cell death and ROS generation against oxidative stress than AV. Thus, CS was selected for subsequent experiments. Ethyl acetate (EA), hexane, butanol, aqueous, and chloroform extracts from CS significantly inhibited cell death and ROS generation in SK-N-SH cells induced by oxidative stress. EA extract from CS (CS-EA) showed the highest DPPH radical-scavenging activity, intra-cellular ROS-scavenging activity, and neuroprotective effects. CS-EA attenuated apoptosis signal-regulating p38 activation by inhibiting phosphorylation. The findings suggest that CS-EA protects neuronal cells through antioxidant activity and inhibition of phosphorylation of p38 in brain neural cells.

신경계 질환은 산화적 스트레스에 의한 신경세포 손상에 의해 발생하는 것이 하나의 기전으로 알려져 있다. 본 연구는 $H_2O_2$에 의해 유도된 산화적 스트레스에 대항하여 곤드레(Cirsium setidens, CS), 누룩치(Pleurospermum kamtschaticumin, PK) 그리고 산마늘(Allium victorials, AV)의 뇌신경 보호 효과 및 그 기전에 대한 것이다. CS와 AV 처리는 대조군과 비교하여 $400{\mu}g/mL$까지 인간의 신경세포주인 SK-N-SH 세포에 대해 세포독성이 없었다. 산화적 유도자인 $H_2O_2$를 SK-N-SH 세포에 처리하였을 때 세포사멸 및 활성산소종(ROS) 생산이 현저하게 증가하였으나 CS 또는 AV 처리에 의해 산화적 스트레스에 의해 증가된 세포사멸과 ROS 생산이 현저하게 감소하였다. 실험한 산채 중에 CS와 PK가 AV보다 더 강한 DPPH 라디칼 소거 작용이 있었으나 PK는 대조군과 비교하여 SK-N-SH 세포를 사멸시키는 강한 세포독성을 가지고 있었다. CS는 AV보다 산화적 스트레스에 대항하여 세포사멸 및 ROS 생성에 더 높은 저해적 영향력을 보여주었다. 따라서 계속되는 실험에는 CS를 사용하였다. CS의 순차적 용매 분획물들인 헥산, 클로로포름, 에틸아세테이트, 부탄올 및 물 분획물들(CS-HE, CS-CH, CS-EA, CS-BU, CS-AQ)은 산화적 스트레스에 대항하여 SK-N-SH 세포사멸 및 세포내 ROS 생성을 억제하였다. CS-EA는 5개의 분획물들 중 가장 강한 DPPH 라디칼 소거작용 및 세포내 ROS 소거 활성을 가지고 있었고, 가장 강한 뇌신경세포 보호 효과를 가지고 있었다. CS-EA는 산화적 스트레스에 의해 증가된 세포자멸사(apoptosis)의 신호전달 경로에 관여하는 p38의 인산화를 저해함으로써 활성화되는 것을 약화시켰다. 이 결과들은 CS-EA가 뇌신경세포에서 항산화 효과 및 p38 인산화 억제에 의한 뇌신경 보호 효과를 나타낼 것이라 제안하였다.

Keywords

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