Journal of Nutrition and Health

  • 제48권5호
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  • Pages.390-397
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  • 2015
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  • 2288-3886(pISSN)
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  • 2288-3959(eISSN)
한국영양학회 (The Korean Nutrition Society)
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만성 알코올과 철분의 과잉 섭취가 흰쥐의 간 세포 미토콘드리아 DNA 손상에 미치는 영향

Effects of chronic alcohol and excessive iron intake on mitochondrial DNA damage in the rat liver

  • 박정은 (경희대학교 생활과학대학 식품영양학과) ;
  • 이정란 (경희대학교 생활과학대학 식품영양학과) ;
  • 정자용 (경희대학교 생활과학대학 식품영양학과)
  • Park, Jung-Eun (Department of Food and Nutrition, Kyung Hee University) ;
  • Lee, Jeong-Ran (Department of Food and Nutrition, Kyung Hee University) ;
  • Chung, Jayong (Department of Food and Nutrition, Kyung Hee University)
  • 투고 : 2015.08.18
  • 심사 : 2015.10.08
  • 발행 : 2015.10.31
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초록

본 연구에서는 Sprague-Dawley 종 랫트 수컷을 대조군, EtOH군, Fe군, EtOH + Fe군으로 나누어, 알코올과 철분을 액상 사료로 8주간 공급한 후, 간 조직과 간 세포 mtDNA의 손상 정도를 알아보았다. EtOH + Fe군은 대조군, EtOH군, Fe군의 다른 세 군에 비해 혈청 ALT와 혈청 AST 수치가 가장 유의적으로 높았으며, 간 조직 검사의 결과에서도 다수의 지방구, 염증성 세포 침입 및 조직의 괴사가 관찰되는 등 가장 심한 간 손상이 확인되었다. DNA 손상 여부를 긴 영역 PCR을 사용하여 분석한 결과, 만성적인 알코올과 철분에 의한 노출은 간 세포의 mtDNA 손상을 유발하는 것으로 나타났으며, 핵 DNA에는 영향을 미치지 않았다. 또한 미토콘드리아의 호흡에 관여하는 Cox1과 Nd4 유전자 발현 정도를 real-time PCR으로 분석한 결과, 알코올 또는 철분은 간 세포의 Cox1 mRNA와 Nd4 mRNA 수준을 유의적으로 낮추는 것으로 나타났다. 이상의 결과는 만성 알코올 또는 과잉의 철분에 의한 간 손상에 mtDNA 손상 및 미토콘드리아 기능 저하가 관여함을 제시한다.

Purpose: In this study, we investigated the effects of chronic alcohol and excessive iron intake on mitochondrial DNA (mtDNA) damage and the progression of alcoholic liver injury in rats. Methods: Twenty-four Sprague-Dawley male rats were divided into four groups (Control, EtOH, Fe, and EtOH + Fe), and fed either control or ethanol (36% of total calories) liquid diet with or without 0.6% carbonyl iron for eight weeks. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, liver malondialdehyde concentrations were measured by colorimetric assays. Liver histopathology was examined by Hematoxylin-eosin staining of the fixed liver tissues. The integrity of the hepatic mtDNA and nuclear DNA was measured by long-range PCR. The gene expression levels of cytochrome c oxidase subunit 1 (Cox1) and NADH dehydrogenase subunit 4 (Nd4) were examined by real-time PCR. Results: Serum ALT and AST activities were significantly higher in the EtOH+Fe group, as compared to the Control group. Similarly, among four groups, liver histology showed the most severe lipid accumulation, inflammation, and necrosis in the EtOH + Fe group. PCR amplification of near-full-length (15.9 kb) mtDNA showed more than 50% loss of full-length product in the liver of the EtOH + Fe group, whereas amounts of PCR products of a nuclear DNA were unaffected. In addition, the changes in the mtDNA integrity showed correlation with reductions in the mRNA levels of mitochondrial gene Cox1 and Nd4. Conclusion: Our data suggested that the liver injury associated with excessive iron and alcohol intake involved mtDNA damage and corresponding mitochondrial dysfunction.

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피인용 문헌

  1. 방사선에 의한 미토콘드리아 손상의 형태학적 고찰 vol.43, pp.1, 2020, https://doi.org/10.17946/jrst.2020.43.1.29