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Ethanol Extract of Ganoderma lucidum Augments Cellular Anti-oxidant Defense through Activation of Nrf2/HO-1

  • Lee, Yoo-hwan (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine) ;
  • Kim, Jung-hee (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine) ;
  • Song, Choon-ho (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine) ;
  • Jang, Kyung-jeon (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine) ;
  • kim, Cheol-hong (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine) ;
  • Kang, Ji-Sook (Anti-Aging Research Center, Dong-Eui University) ;
  • Choi, Yung-hyun (Anti-Aging Research Center, Dong-Eui University) ;
  • Yoon, Hyun-Min (Departments of Acupuncture and Moxibustion, Dong-Eui University College of Korean Medicine)
  • Received : 2016.01.24
  • Accepted : 2016.02.17
  • Published : 2016.03.31

Abstract

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)

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