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

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Antioxidant Activity and Protective Effects of Extracts from Chrysanthemum boreale on t-BHP Induced Oxidative Stress in Chang Cells

산국대 추출물의 항산화 활성 및 간세포 보호 효과

  • Received : 2013.09.11
  • Accepted : 2013.12.23
  • Published : 2014.01.31
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Abstract

The aim of this study was to evaluate the antioxidant activity and protective effect of extracts from the stems and leaves of Chrysanthemum boreale (CBSL) on t-BHP induced oxidative stress in human liver cells (Chang cells). Antioxidant activities in the extracts were determined for various radical scavenging activities including ferric reducing antioxidant power, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, and oxygen radical absorbance capacity (ORAC). CBSL showed a very good scavenging effect of DPPH radical ($IC_{50}$ $0.009{\pm}0.002$ mg/mL), alkyl radical ($IC_{50}$ $0.004{\pm}0.001$ mg/mL), and hydroxyl radical ($IC_{50}$ $6.742{\pm}0.152$ mg/mL). CBSL also showed a strong antioxidant effect in the ORAC assay. In the MTT assay on human liver cells (Chang cells), the extracts showed protective effects by increasing cell viability, decreasing ROS, and restoring mitochondria membrane potential upon t-BHP induced oxidative stress. Our findings suggest that CBSL extracts are a potential therapeutic with protective antioxidant effects upon oxidative stress.

본 연구는 산국대 추출물의 항산화 활성을 탐색하고자 산국대 추출물에 포함된 총 폴리페놀 함량, 총 플라보노이드 함량, 전자스핀공명기기를 이용한 DPPH, alkyl 라디칼 소거능, ABTS를 이용한 라디칼 소거 활성, FRAP를 이용한 총 항산화능 및 ORAC를 측정하였다. 또한 세포 독성 및 간세포 보호 효능 실험을 수행하였다. 산국대 추출물의 총 플라보노이드 함량은 $4.41{\pm}0.04$ mg GAE/g, 총 폴리페놀 함량은 $4.75{\pm}0.03$ mg CE/g으로 나타났다. Hydroxyl 소거 활성은 대조군으로 사용한 비타민 C보다 낮았지만, DPPH 라디칼 소거능이나 alkyl 라디칼 소거능은 비타민 C와 비슷하거나 오히려 더 높은 활성을 나타내었다. 또한 ABTS를 이용한 라디칼 소거 활성과 FRAP를 이용한 총 항산화능 측정을 통한 항산화 활성을 평가한 결과에서도 산국대 추출물이 항산화 효과를 가지고 있음이 확인되었다. 게다가 DNA strand break 평가에서 산국대 추출물이 농도 의존적인 DNA 보호효과를 가지고 있음을 보여주었다. 세포 독성을 살펴보기 위하여 정상 간세포(human liver cells, Chang cells)를 이용하여 MTT assay를 수행한 결과 세포의 생존율은 0.2mg/mL의 농도까지는 전혀 독성을 나타내지 않았고, 간세포 보호 효능 실험에서는 샘플을 한 시간 동안 전처리했을 때 t-BHP로 유도한 산화적 스트레스에 대해 농도 의존적으로 생존율이 증가하는 것을 확인하였다. 세포내 활성산소종을 측정한 결과와 미토콘드리아 막 전위차를 측정한 결과에서 산화적 스트레스에 대한 간세포 보호 효과를 나타내었다. 이는 산국대 추출물이 가진 항산화 활성이 세포내 활성산소종의 소거를 통한 산화스트레스를 억제하여 세포를 보호하는 효과가 있는 것으로 판단된다. 이상의 결과로 기존 산국에 관한 연구는 꽃에 관한 연구가 주를 이루고 있고 줄기와 잎에 관한 연구가 부족한 실정이기에, 산국대를 기능성 소재로 활용할 때 기초자료로 활용할 뿐 아니라 꽃보다 수확량이 많으므로 경제적인 소재활용이 될 수 있을 것으로 판단된다.

Keywords

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