Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Effects of Acanthopanax senticosus Water Extract on Glucose-Regulating Mechanisms in HepG2 Cells

가시오갈피 물 추출물이 간세포에서 포도당 이용 대사에 미치는 영향

  • Kim, Dae-Jung (Well-being Bioproducts RIC, Kangwon National University) ;
  • Kang, Yun Hwan (National Development Institute of Korean Medicine) ;
  • Kim, Kyoung Kon (Department of Bio-Health Technology, Kangwon National University) ;
  • Kim, Tae Woo (Well-being Bioproducts RIC, Kangwon National University) ;
  • Park, Jae Bong (Department of Biochemistry, Hallym University College of Medicine) ;
  • Choe, Myeon (Well-being Bioproducts RIC, Kangwon National University)
  • 김대중 (강원대학교 웰빙특산물산업화지역혁신센터) ;
  • 강윤환 (한약진흥재단) ;
  • 김경곤 (강원대학교 생명건강공학과) ;
  • 김태우 (강원대학교 웰빙특산물산업화지역혁신센터) ;
  • 박재봉 (한림대학교 의과대학 생화학교실) ;
  • 최면 (강원대학교 웰빙특산물산업화지역혁신센터)
  • Received : 2016.12.14
  • Accepted : 2017.04.17
  • Published : 2017.05.31
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Abstract

This study aimed to investigate glucose uptake mechanisms and metabolic mechanisms for absorbed glucose in HepG2 cells treated with Acanthopanax senticosus water extract (ASW). A colorimetric assay kit was used to measure polyphenol content, glucokinase (GK) activity, glucose uptake, glucose consumption in cell culture medium, and glycogen content. RT-PCR and western blotting were performed to examine changes in the expression levels of glucose transporter 2 (GLUT2), hepatocyte nuclear factor $1{\alpha}$ ($HNF-1{\alpha}$), phosphatidylinositol 3-kinase (PI3k), protein kinase B (Akt), phospho-AMP-activated protein kinase (AMPK), phosphoenolpyruvate carboxykinase, GK, and glycogen synthase kinase $3{\beta}$ ($GSK3{\beta}$). Increased glucose uptake upon ASW treatment was confirmed to result from increased expression of $HNF-1{\alpha}$, which is one of the transcription factors acting on the GLUT2 promoter. From the measurements of GK activity, we observed that ASW had an effect on glucose phosphorylation, and we also confirmed that increased AMPK phosphorylation promoted glycolysis and suppressed gluconeogenesis. We confirmed that the increase in glycogen upon ASW treatment was induced by activation of Akt by PI3k, followed by phosphorylation of $GSK3{\beta}$. This study demonstrates that ASW activates glucose metabolic mechanisms in liver cells and is therefore a potential candidate to alleviate diabetes.

본 연구에서는 가시오갈피 물 추출물(ASW)를 이용하여 아직 시도된 바가 없는 HepG2 세포 내 포도당 유입과정 및 glucokinase(GK) 활성을 통한 포도당 이용대사 실험을 수행하였다. 포도당의 세포 내 유입은 GLUT2의 transcription factor들 중 하나인 $HNF-1{\alpha}$의 활성화로 GLUT2의 유전자 발현이 증가하여 이루어지는 것을 확인하였다. GK 활성 측정 결과 ASW가 GK를 활성화하여 포도당의 인산화에 영향을 주는 것을 확인하였고 AMP-activated protein kinase의 인산화 증가로 glycolysis에 관여하는 효소인 GK의 단백질 발현은 증가하고, gluconeogenesis에 관여하는 phosphoenolpyruvate carboxykinase의 단백질 발현은 감소하는 것을 확인하였다. 그리고 인산화된 포도당이 glycogen으로 전환 저장되는 메커니즘을 pPI3k-pAkt-pGSK-$3{\beta}$의 단계별 단백질 발현을 확인함으로써 검증하였으며, glycogen 함량 측정을 통해 확인하였다. 본 연구를 통해 ASW가 다양한 메커니즘에 작용하여 당뇨의 예방 및 개선에 활용할 수 있는 잠재적 소재임을 확인하였고, 이는 ASW가 천연 기능성 소재로서의 개발가치가 높음을 시사한다.

Keywords

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