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Protective Effects of Sasa Borealis Leaves Extract on High Glucose-Induced Oxidative Stress in Human Umbilical Vein Endothelial Cells

혈관내피세포에서 고농도 포도당으로 유도된 산화스트레스에 대한 조릿대잎 추출물의 보호효과

  • Hwang, Ji-Young (Dept. of Food Science and Nutrition, Pusan National University) ;
  • Han, Ji-Sook (Dept. of Food Science and Nutrition, Pusan National University)
  • Received : 2010.09.06
  • Accepted : 2010.11.29
  • Published : 2010.12.31

Abstract

This study was designed to investigate the protective effects of Sasa borealis leaves on high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). Freeze-dried Sasa borealis leaves were extracted with 70% methanol and followed by a sequential fractionation with dicholoromethan, ethyl acetate, butanol and water. The ethyl acetate fraction from Sasa borealis leaves extract (ESLE) was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. Exposure of HUVECs to 30 mM high glucose for 48 hr resulted in a significant (p<0.05) decrease in cell viability, glutathion (GSH) concentration, activities of antioxidant enzymes including superoxide dimutase (SOD), glutathion peroxidase (GSH-px) and catalase, and a significant (p<0.05) increase in intracellular ROS and lipid peroxidation formation in comparison to the cells treated with 5.5 mM glucose. ESLE treatment decreased intracellular ROS and lipid peroxidation formation and increased cell viability, GSH concentration and expressions of SOD and catalase in HUVECs. These results suggest that ESLE may be able to protect HUVECs from high glucose-induced oxidative stress, partially through the antioxidative defense systems.

혈관내피세포는 만성적인 고혈당 상태에 노출되면, 반응성 산소기의 급격한 증가로 인해 산화스트레스를 일으키게 된다. 이를 보호하기 위해 사용되는 항산화제는 혈관내피세포의 생존율을 높이며 2차적인 질병의 유발을 감소 및 완화시켜서 당뇨합병증에 도움이 되는 것으로 알려져 있는데, 본 연구에서는 항산화 효과가 있는 것으로 알려진 조릿대잎 추출물을 이용하여 고농도의 포도당으로 유도된 산화스트레스에 대한 혈관내피세포의 보호효과를 조사하고자 하였다. 즉, 30 mM 포도당에 노출되어 산화적 스트레스가 유발된 HUVECs에 조릿대잎 추출물 ethyl acetate(ESLE) 층을 분주한 결과, 농도 의존적으로 세포 생존율이 증가하였고, 활성산소종 수준과 지질과산화물 가는 ESLE 첨가에 의해 유의적으로 감소하는 것을 확인할 수 있었다. 또한 고농도 포도당으로 GSH 함량과 SOD, GSH-px, catalase 등과 같은 항산화효소의 활성이 감소된 HUVECs에 ESLE를 분주하였을 때 이들의 함량 및 활성이 유의적으로 증가되었으며, western blotting을 통해 ESLE 첨가가 SOD와 catalase 발현을 증가시킴을 확인할 수 있었다. 따라서 ESLE는 혈관내피세포에서 고농도 포도당으로 인해 유발된 산화적 스트레스에 대해 세포를 보호하는 효과가 있는 것으로 나타났으며, 이는 ESLE가 세포내 활성산소종을 감소시키고 항산화효소계의 활성을 증가시킴으로써 산화스트레스를 감소시킨 결과에 기인하는 것으로 사료되었다.

Keywords

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