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배양 신경세포의 저산소증모델에서 대황 물추출액의 항산화 및 사립체막전위 소실 억제 효능

Water Extract of Rhei Rhizoma Prevent Production of Reactive Oxygen Species and Loss of Mitochondrial Membrane Potential in a Hypoxia Model of Cultured Neurons

  • 이현숙 (동국대학교 의과대학 해부학교실) ;
  • 문일수 (동국대학교 의과대학 해부학교실)
  • Lee, Hyun-Sook (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Moon, Il-Soo (Department of Anatomy, College of Medicine, Dongguk University)
  • 발행 : 2008.12.30

초록

대황(Rhei Rhizoma; RR, 대황(大黃))은 Rheum officinale Baill.와 Rheum palmatum L. (Polygonaceae)의 땅속부분 (rhizome 및 root)으로 남아시아의 민속의학에서 간 및 신장의 손상을 치료하는데 널리 이용되고 있다. 본 연구에서는 배양한 흰쥐 해마신경세포의 저산소증모델을 이용하여 대황의 물추출물이 신경세포사를 억제하는 효능에 대하여 조사하였다. 배양 10일(DIV10)에 RR을 배양액에 첨가하고 DIV13일에 생존율을 조사한 결과 10 ${\mu}g$/ml 농도까지는 세포독성이 없었으며, 정상산소 환경에서 2.5 ${\mu}g$/ml의 농도에서 세포생존율을 높이는 것으로 나타났다. 또한 배지에 대황을 첨가한 경우 DIV13일에 저산소증을 유도한 후 5일째에 세포생존율을 조사한 결과 대조군에 비하여 매우 유의하게 생존율을 증가시켰다. $H_2DCF$ 염색 결과 대황은 활성산소(ROS)의 생성을 유의하게 감소시킴과, JC-1 염색 결과 사립체 막전위의 소실을 유의하게 억제함을 알 수 있었다. 이러한 결과들은 대황 물추출물이 활성산소를 효과적으로 제거하고, 세포의 에너지 생성을 보전함으로서 세포사를 억제할 수 있음을 보여주며, 향후 뇌신경세포의 건강에 유용하게 이용될 수 있음을 시사한다.

Rhei Rhizoma (RR; 大黃) consists of the underground parts (rhizome and root) of Rheum officinale Baill. and Rheum palmatum L. (Polygonaceae), and is widely used in Southeast Asian folk medicine to alleviate liver and kidney damages. In this study, we investigated into the efficacy and mechanism of RR water extract in supporting neuronal survival in a hypoxia model of cultured rat hippocampal neurons. RR exhibited no cytotoxicity up to 10 ${\mu}g$/ml and exhibited neurosupportive effects at 2.5 ${\mu}g$/ml in normoxia. When RR was added to the culture media on 10 days in vitro (DIV10) and given a hypoxic shock (2% $O_2$/5% $CO_2$, 3 hr, $37^{\circ}C$) on DIV13, RR exhibited neuroprotective effects on 5 days post-shock. $H_2DCF$ stainings indicated that RR effectively prevents ROS production in both normoxia and hypoxia. JC-1 stainings showed that RR prevents dissipation of MMP in hypoxia. These results indicate that RR protects neurons by suppressing ROS production and MMP loss.

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참고문헌

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