Clinical and Experimental Pediatrics

The Korean Pediatric Society (대한소아청소년과학회)
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Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats

신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과

  • Shin, Jin Young (Department of Pediatrics, School of Medicine, Catholic University of Daegu) ;
  • Seo, Min Ae (Department of Pediatrics, School of Medicine, Catholic University of Daegu) ;
  • Choi, Eun Jin (Department of Pediatrics, School of Medicine, Catholic University of Daegu) ;
  • Kim, Jin Kyung (Department of Pediatrics, School of Medicine, Catholic University of Daegu) ;
  • Seo, Eok Su (Department of Ophthalmology, Dongguk University College of Medicine) ;
  • Lee, Jun Hwa (Department of Pediatrics, Masan Samsung Hospital, School of Medicine, Sungkyunkwan University) ;
  • Chung, Hai Lee (Department of Pediatrics, School of Medicine, Catholic University of Daegu) ;
  • Kim, Woo Taek (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
  • 신진영 (대구가톨릭대학교 의과대학 소아과학교실) ;
  • 서민애 (대구가톨릭대학교 의과대학 소아과학교실) ;
  • 최은진 (대구가톨릭대학교 의과대학 소아과학교실) ;
  • 김진경 (대구가톨릭대학교 의과대학 소아과학교실) ;
  • 서억수 (동국대학교 의과대학 안과학교실) ;
  • 이준화 (성균관대학교 의과대학 마산삼성병원 소아청소년과) ;
  • 정혜리 (대구가톨릭대학교 의과대학 소아과학교실) ;
  • 김우택 (대구가톨릭대학교 의과대학 소아과학교실)
  • Received : 2008.06.02
  • Accepted : 2008.09.01
  • Published : 2008.10.15
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Abstract

Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

목 적: Resveratrol은 주로 포도나무의 과실이나 잎 부위에서 추출되는 성분으로, 주로 심질환에서 암 예방 효과, 항염증 효과, 항산화 효과의 기능이 밝혀지고 있다. 최근 성인에 대한 신경보호 효과가 있는 것으로 알려졌지만. 신생아에서 연구는 아직까지 없다. 그래서 본 연구에서는 resveratrol이 신생 백서의 저 산소 허혈 뇌손상에서 신경보호 효과가 있는지를 알아보고자 실험하였다. 방 법: 재태기간 18일된 태아 백서의 대뇌피질 세포를 배양하여 1% $O_2$ 배양기에서 저 산소 상태로 뇌세포손상을 유도하여 저 산소군, 저 산소 30분 전 resveratrol 투여군 (1, 10, $30{\mu}g/mL$)으로 나누어 정상 산소군과 비교하였다. 또한, 동물 모델에서는 생후 7일된 백서의 좌측 총 경동맥을 결찰한 후 저 산소 (8% $O_2$) 상태로 2.5시간 노출시켜서 저 산소 허혈 뇌 손상을 유발하였고, 뇌손상 전후 30분에 resveratrol을 체중 kg당 30 mg을 복막내로 투여하였다. 세포사멸사의 관련을 알아보기 위해 Bcl-2, Bax, caspase-3 primer를 이용한 실시간 중합효소연쇄반응과 동일 항체를 이용한 Western blotting을 시행하였다. 결 과: 태아 백서 뇌세포 배양 실험에서 저 산소군의 경우 Bcl-2의 발현이 정상 산소군에 비해 감소하였고, Bax의 발현과 caspase-3의 발현, 그리고 Bax/Bcl-2의 비율은 증가하였다. Reaveratrol을 투여한 실험군의 경우에서는 Bcl-2 발현은 증가하였고, Bax의 발현과 caspase-3의 발현, Bax/Bcl-2의 비율은 저 산소군에 비하여 감소하는 결과를 보였다. 또한 이는 저 산소 허혈 뇌손상 동물 모델에서도 같은 결과를 보였다. 결론: 본 연구에서 resveratrol은 주산기 저 산소 허혈 뇌손상에서 세포사멸사 작용의 억제를 통하여 신경보호 역할을 하는 것을 알 수 있었다.

Keywords

Acknowledgement

Supported by : Catholic University of Daegu

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