Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Global DNA Methylation Patterns and Gene Expression Associated with Obesity-Susceptibility in Offspring of Pregnant Sprague-Dawley Rats Exposed to BDE-47 and BDE-209

임신 중 BDE-47 및 BDE-209에 노출된 어미와 새끼 Sprague-Dawley 랫드의 Global DNA 메틸화 양상과 비만 감수성과 연관된 유전자 발현

  • Park, Byeong-Min (Department of Laboratory Medicine, Research Institute and Hospital, National Cancer Center) ;
  • Yoon, Ok-Jin (Department of Laboratory Medicine, Research Institute and Hospital, National Cancer Center) ;
  • Lee, Do-Hoon (Department of Laboratory Medicine, Research Institute and Hospital, National Cancer Center)
  • 박병민 (국립암센터 진단검사의학과) ;
  • 윤옥진 (국립암센터 진단검사의학과) ;
  • 이도훈 (국립암센터 진단검사의학과)
  • Received : 2016.12.27
  • Accepted : 2017.02.14
  • Published : 2017.03.31
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Abstract

Persistent organic pollutants (POPs) can affect epigenetic mechanisms and obesity development. Polybrominated diphenyl ethers (PBDEs)-widely used to make flames-are one of the important POPs. Prenatal exposure to endocrine disrupting chemicals (EDCs), such as POPs, may affect global DNA methylation in long interspersed nuclear elements (LINE-1), increasing the risk of obesity later in life. Therefore, pregnant Sprague-Dawley (SD) rats were used to elucidate whether BDE-47 and BDE-209 transferred through placenta and breast milk cause epigenetic changes in LINE-1 and increase genetic susceptibility to obesity as obesogen during the developmental periods. Global DNA methylation in LINE-1 and gene expression related to obesity were measured in dams and offspring, using a methylation-sensitive high resolution melting analysis (MS-HRM) and direct bisulfite sequencing and quantitative real time polymerase chain reaction (qPCR), respectively. The results of MS-HRM showed global DNA hypomethylation patterns in LINE-1 of exposed offspring (2 of total 4) at PND 4, but bisulfite sequencing showed no difference in both the exposed and non-exposed groups. Gene expression in dams related to ${\beta}$-oxidation pathway and those related to adipokines showed different patterns between the two groups. On the contrary, gene expressions of offspring showed a similar pattern. Gene expressions related to ${\beta}$-oxidation pathway and obesity were significantly increased when compared with 'at birth', but not $PPAR-{\alpha}$. In conclusion, this study demonstrated the possibility that co-exposure to BDE-47 and BDE-209-via the placenta and breast milk-may affect epigenetic changes and modulate gene expression levels related to obesity.

잔류성 유기 오염 물질은 후성학적 메커니즘과 비만의 발달에 영향을 줄 수가 있다. 폴리브롬화 디페닐 에테르는 주요한 잔류성 유기 오염 물질 중 하나이며, 난연제로 널리 쓰인다. 출생전 잔류성 유기 오염 물질과 같은 내분비교란물질에 노출시 LINE-1 (long interspersed nuclear elements)의 global DNA 메틸화와 비만 위험도의 증가에 영향을 미칠 수 있다. 따라서, 이번 연구는 임신한 스프라그-돌리 백서를 이용하여 태반과 모유를 통하여 전달된 BDE-47, BDE-209가 LINE-1에서의 후성학적인 변화와 obesogen으로서 발달과정에 따른 유전적 비만 감수성의 증가에 영향을 줄 수 있는지에 대해서 보고자 하였다. 어미와 새끼에서 LINE-1의 광범위 DNA 메틸화와 비만과 관련된 유전자 발현은 methylation-sensitive high resolution melting analysis (MS-HRM), direct bisulfite sequencing와 quantitative real time polymerase chain reaction (qPCR)을 사용하여 각각 분석하였다. MS-HRM 결과는 출생 후 4일의 노출군 새끼에서 (4마리 중 2마리) LINE-1의 광범위 DNA 저메틸화 양상을 보여주었지만, bisulfite sequencing은 노출군과 비노출군에서 차이가 없었다. ${\beta}$-산화 경로와 adipokines과 관련된 어미의 유전자 발현은 두 그룹간 차이를 보였다. 반면에, 새끼의 유전자 발현은 비슷한 양상을 나타내었다. ${\beta}$-산화 경로와 비만과 관련된 유전자 발현 중 $PPAR-{\alpha}$를 제외하고는 출생 시에 유의하게 증가하였다. 결론적으로, 이번 연구는 BDE-47, BDE-209의 동시 노출이 태반과 모유를 통해서 새끼에서의 후성학적인 변화와 비만과 관련된 유전자 발현 변화에 영향을 미칠 수 있는 것을 보여주었다.

Keywords

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