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PC12 세포에서 생물학적 활성에 미치는 원지 열수 추출물의 효능

Effect of Polygala radix Hot Water Extract on Biological Activity in PC12 Cells

  • 투고 : 2013.07.29
  • 심사 : 2013.08.27
  • 발행 : 2013.08.30

초록

원지의 뿌리는 기억력을 향상시키는 동양의 전통 약재로 널리 알려져 있다. 그러나 그 작용기전은 아직까지 해명되지 않았다. 본 연구에서는 항산화 효과 뿐만 아니라 신경 세포 에서 파생된 PC12 세포의 acetylcholinesterase(AchE) 활성과 연관된 인지능에 대한 원지열수추출물(PRHWE)의 효과를 in vitro 및 살아 있는 세포에서 조사하였다. 먼저 MTT assay을 이용한 세포생존에 대한 연구에서 PRHWE는 0.1% 이하의 농도에서 세포독성이 없는 것으로 나타났다. PRHWE는 DPPH radical, hydrogen peroxide 및 superoxide의 소거활성과 환원력이 농도에 비례하여 증가시킨다는 것이 관찰되었다. 특히 PRHWE는 hydroxyl radical에 의하여 유발되는 DNA의 산화에 대한 보호효과를 나타내었다. 부가적으로 그것은 신경성세포에서 nitric oxide의 생성을 억제하였다. 더욱이 AchE 활성은 PRHWE의 농도에 비례하여 감소하였다. 뿐만 아니라, PRHWE는 PC12 세포에서 SOD-1과 NOS-2 발현수준을 증가 시킨다는 것이 발견되었다. 더욱이 reporter gene assay를 이용한 실험에서 p53과 NF-${\kappa}B$의 전사활성이 PRHWE의 존재 하에서 감소했다. 그러므로 이러한 결과들은 PRHWE가 신경성세포에서 항산화 활성 및 신경 세포 보호 효과가 있다는 것을 증명하였고, 이는 PRHWE가 사람의 건강을 위한 치료제로 큰 잠재성을 가지고 있다는 것을 시사한다.

The root of Polygala radix has been widely known as an oriental traditional medicinal stuff that improves memory. However, its mechanism of action remains unclear. In this study, the effect of Polygala radix hot water extracts (PRHWE) on cognitive function related to the activity of acetylcholinesterase (AchE) derived from neural cells (PC12) in addition to antioxidant activity was examined both in a cell-free system and live cells. First, in the study on cell viability using an MTT assay, PRHWE did not exhibit any cell toxicity at 0.1% (w/v) or below. It also was observed that PRHWE increased the scavenging activity of DPPH radical, hydrogen peroxide and superoxide, reducing power in a dose-dependent manner. In particular, PRHWE had a protective effect on DNA oxidation induced by hydroxyl radicals. Additionally, it inhibited the production of inducible nitric oxide in neuronal cells. Furthermore, the AchE activity decreased with increasing concentrations. In addition, PRHWE increased the expression level of SOD-1 and NOS-2 in PC12 cells. Moreover, the transcriptional activities of p53 and NF-${\kappa}B$ were reduced in the presence of PRHWE in an experiment using a reporter gene assay. Therefore, these results prove that PRHE has antioxidative and protective effects on neuronal cells, suggesting that it may have great potential as a therapeutic agent for human health.

키워드

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