Protective Effect of Methanolic Extracts from Dendrobium nobile Lindl. on $H_{2}O_{2}$-induced Neurotoxicity in PC12 cells

석곡 MeOH 추출물이 $H_{2}O_{2}$에 의한 신경세포 보호효과에 미치는 영향

  • Yoon, Mi-Young (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Kim, Ju-Young (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Hwang, Ji-Hwan (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Cha, Mi-Ran (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Lee, Mi-Ra (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Jo, Kyung-Jin (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Park, Hae-Ryong (Department of Food Science and Biotechnology, Kyungnam University)
  • 윤미영 (경남대학교 식품생명학과) ;
  • 김주영 (경남대학교 식품생명학과) ;
  • 황지환 (경남대학교 식품생명학과) ;
  • 차미란 (경남대학교 식품생명학과) ;
  • 이미라 (경남대학교 식품생명학과) ;
  • 조경진 (경남대학교 식품생명학과) ;
  • 박해룡 (경남대학교 식품생명학과)
  • Published : 2007.03.31

Abstract

The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.

본 연구에서는 석곡(Dendrobium nobile Lindl.) 추출물에 대한 신경세포 보호 효과를 확인하기 위하여 $H_{2}O_{2}$에 의해 유도된 스트레스 상태의 PC12 세포주에서 MTT-dye reduction assay와 LDH release assay를 이용하였다. MTT reduction assay 결과, 스트레스 상태에서는 47%의 생존율을 보인데 반하여 석곡 추출물 50${\mu}$g/ml 농도로 처리하였을 때 99.5%의 높은 세포 생존율을 확인할 수 있었다. 이 결과는 LDH release assay 에서도 일치하는 결과를 확인하였다. 그리고 광학 현미경을 이용한 형태학적 변화를 관찰한 결과에서도 신경돌기의 출현 유도를 통한 신경세포 생존을 확인 할 수 있었다. 또한 핵의 변화에 미치는 영향을 관찰한 결과 정상 세포의 핵은 타원형의 온전한 핵 모양을 나타낸 반면 산화적 손상을 입은 세포는 apoptotic body가 핵 주변에 나타나는 전형적인 apoptosis를 나타내었고 석곡 추출물을 처리한 결과 핵의 condersation현상과 fragmentation이 현저히 감소함을 확인 할 수 있었다. 그리고 $H_{2}O_{2}$에 의한 capase-3의 활성은 석곡 추출물을 50${\mu}$g/ml 처리 후 약 1.1배 이하로 감소함을 보였다.

Keywords

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