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청미래덩굴 잎 물추출물이 처리된 HepG2 세포에서의 포도당흡수기전 연구

Study of the mechanisms underlying increased glucose absorption in Smilax china L. leaf extract-treated HepG2 cells

  • 강윤환 (강원대학교 강원웰빙특산물산업화지역혁신센터) ;
  • 김대중 (강원대학교 강원웰빙특산물산업화지역혁신센터) ;
  • 김경곤 (강원대학교 생명건강공학과) ;
  • 이성미 (강원대학교 생명건강공학과) ;
  • 최면 (강원대학교 강원웰빙특산물산업화지역혁신센터)
  • Kang, Yun Hwan (Well-being Bioproducts RIC, Kangwon National University) ;
  • Kim, Dae Jung (Well-being Bioproducts RIC, Kangwon National University) ;
  • Kim, Kyoung Kon (Department of Bio-Health Technology, Kangwon National University) ;
  • Lee, Sung Mee (Department of Bio-Health Technology, Kangwon National University) ;
  • Choe, Myeon (Well-being Bioproducts RIC, Kangwon National University)
  • 투고 : 2014.02.26
  • 심사 : 2014.04.22
  • 발행 : 2014.06.30

초록

본 연구에는 SCLE를 이용하여 시도된 바가 없는 glucose uptake 유도 실험을 수행하여 HepG2 세포에서 포도당흡수가 증가함을 확인하였다. 또한 이런 포도당의 흡수는 HNF-$1{\alpha}$라는 transcription factor의 활성화를 통해 GLUT-2의 발현을 증가시키기 때문인 것을 실험적으로 증명하였다. 뿐만 아니라 Bacillus stearothermophilus 유래의 GK를 이용하여 활성을 측정한 결과 SCLE가 직접적으로 GK를 활성화하여 포도당의 인산화에 영향을 주는 것을 확인할 수 있었으며 그 실험결과들은 Fig. 8에서 도식화 하였다. 본 연구를 통해 SCLE가 ${\alpha}$-glucosidase inhibition 활성에 의한 혈당의 개선과 당뇨예방 효과뿐만 아니라 다양한 세포내 기전을 통해 혈당 및 당뇨의 개선을 유도할 수 있음을 확인하였고, 이는 SCLE가 neutraceuticals 소재로서의 개발가치가 높음을 시사한다.

Purpose: Previous studies have shown that treatment with Smilax china L. leaf extract (SCLE) produces antidiabetic effects due to ${\alpha}$-glucosidase inhibition. In this study, we examined the mechanism underlying these antidiabetic effects by examining glucose uptake in HepG2 cells cultured with SCLE. Methods: Glucose uptake and glucokinase activity were examined using an assay kit. Expression of glucose transporter (GLUT)-2, GLUT-4, and HNF-$1{\alpha}$ was measured by RT-PCR or western blot. Results: Treatment with SCLE resulted in enhanced glucose uptake in HepG2 cells, and this effect was especially pronounced when cells were cultured in an insulin-free medium. SCLE induced an increase in expression of GLUT-2 but not GLUT-4. The increase in the levels of HNF-$1{\alpha}$, a GLUT-2 transcription factor, in total protein extract and nuclear fraction suggest that the effects of SCLE may occur at the level of GLUT-2 transcription. In addition, by measuring the change in glucokinase activity following SCLE treatment, we confirmed that SCLE stimulates glucose utilization by direct activation of this enzyme. Conclusion: These results demonstrate that the potential antidiabetic activity of SCLE is due at least in part to stimulation of glucose uptake and an increase in glucokinase activity, and that SCLE-stimulated glucose uptake is mediated through enhancement of GLUT-2 expression by inducing expression of its transcription factor, HNF-$1{\alpha}$.

키워드

참고문헌

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