Herbal Formula Science (대한한의학방제학회지)

The Korean Medicine Society for the Herbal Formula Study (대한한의학방제학회)
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Study of hepatoprotective effect of Haegan-jeon through activation of nuclear factor erythroid 2-related factor 2 and optimization of herbal composition based on molecular mechanism

Nuclear factor erythroid 2-related factor 2 활성화를 통한 해간전(解肝煎)의 간세포 보호 효능 및 분자기전을 활용한 해간전(解肝煎) 구성 약물의 최적화 연구

  • Kim, Jae Kwang (College of Korean Medicine, Daegu Haany University) ;
  • Jung, Ji Yun (College of Korean Medicine, Daegu Haany University) ;
  • Park, Sang Mi (College of Korean Medicine, Daegu Haany University) ;
  • Park, Chung A (College of Korean Medicine, Daegu Haany University) ;
  • Ku, Sae Kwang (College of Korean Medicine, Daegu Haany University) ;
  • Byun, Sung Hui (College of Korean Medicine, Daegu Haany University) ;
  • Cho, Il Je (College of Korean Medicine, Daegu Haany University) ;
  • Kim, Sang Chan (College of Korean Medicine, Daegu Haany University)
  • 김재광 (대구한의대학교 한의과대학) ;
  • 정지윤 (대구한의대학교 한의과대학) ;
  • 박상미 (대구한의대학교 한의과대학) ;
  • 박정아 (대구한의대학교 한의과대학) ;
  • 구세광 (대구한의대학교 한의과대학) ;
  • 변성희 (대구한의대학교 한의과대학) ;
  • 조일제 (대구한의대학교 한의과대학) ;
  • 김상찬 (대구한의대학교 한의과대학)
  • Received : 2018.07.03
  • Accepted : 2018.07.23
  • Published : 2018.08.31
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Abstract

Objectives : Present study investigated hepatoprotective effect of Haegan-jeon extract (HE) and tried to elucidate molecular mechanism involved. According to molecular mechanism, present study optimized herbal composition of HE (op-HE) and compared in vitro and in vivo hepatoprotective effects of op-HE to HE. Methods : For in vitro experiments, HepG2 cells were exposed to arachidonic acid (AA, $10{\mu}M$) and iron ($5{\mu}M$) for inducing oxidative stress. Cell viability, GSH contents, $H_2O_2$ production, mitochondrial membrane potential, immunoblot and reporter gene assay were performed to investigate cytoprotective effects and responsible molecular mechanisms. For in vivo experiments, hepatoprotective effect of HE and op-HE were assessed on $CCl_4-induced$ liver injury mice model. Results : HE pretreatment prevented AA+iron-mediated hepatocytes apoptosis. In addition, AA+iron-induced mitochondrial dysfunction, $H_2O_2$ production, glutathione depletion were reduced by HE pretreatment. In addition, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation, antioxidant response element (ARE)-driven reporter gene activity, and antioxidant genes expression were increased by HE. Based on reporter gene and MTT assays, we found that op-HE consisting three medicinal herbs also significantly increased transactivation of Nrf2 and reduced the AA+iron-mediated cytotoxicity. Moreover, in $CCl_4-induced$ liver injury mice model, HE-op had an ability to ameliorate $CCl_4-mediated$ increases in serum alanine transferase and aspartate aminotransferase activity, hepatic degeneration, inflammatory cell infiltration, and collagen deposition. Hepatoprotective effects of op-HE were comparable to those of HE. Conclusions : Present study suggests that op-HE as well as HE exhibit hepatoprotective effect against oxidative stress-mediated liver injury via Nrf2 activation.

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References

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