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산화 스트레스에 대한 진심유생환(오자연종환 가감방) 추출물의 세포 독성 보호 효과에 관한 연구

Study of the Cytotoxic Protective Effects of Jinsimyusaeng-hwan (Modified Ojayeonjong-hwan ) Against Oxidative Stress

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Objectives : This study was conducted to evaluate the cytotoxic protective effects of Jinsimyusaeng-hwan (Modified Ojayeonjong-hwan ) against oxidative stress. Methods : 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid)(ABTS) radical scavenging activity and ferric reducing ability of plasma (FRAP) method were used to estimate the antioxidant activity of Jinsimyusaeng-hwan. C2C12 myoblasts were used to reevaluate the antioxidant effects. And apoptosis analysis, mitochondrial membrane potential analysis, measurement of intracellular reactive oxygen species levels were conducted to investigate antioxidant activity of Jinsimyusaeng-hwan. Results : In comparison of DPPH free radical and ABTS cationic radical scavenging activity, it increased as the concentration of water extracts of Jinsimyusaeng-hwan (WEJ) and 70% ethanol extracts of Jinsimyusaenghwan (EEJ) increased. In the results of comparing the total phenol content and reducing power using the FRAP method, extracts with high total phenol content also showed high reducing power. In comparison of the protective effect against H2 O2 -induced oxidative stress in C2C12 myoblasts, WEJ had no significant effect, but the EEJ inhibited H2 O2 -mediated cytotoxicity in a concentration-dependent manner. The cytotoxic protective effect of EEJ against oxidative stress in C2C12 myoblasts was correlated with their inhibitory effects on H2 O2 -induced apoptosis and cell cycle arrest. In H2 O2 -treated C2C12 myoblasts, the apoptosis inhibitory effects of EEJ were associated with suppression of mitochondrial dysfunction and DNA damage. The protective effect of EEJ against H2 O2 -induced oxidative stress in C2C12 myoblasts were directly related to the inhibition of ROS generation. Conclusion : Jinsimyusaeng-hwan extracts have cytotoxic protective effects against oxidative stress, and it was better in 70% ethanol extract than in water extract.

Abstract

Objectives : This study was conducted to evaluate the cytotoxic protective effects of Jinsimyusaeng-hwan (Modified Ojayeonjong-hwan ) against oxidative stress. Methods : 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid)(ABTS) radical scavenging activity and ferric reducing ability of plasma (FRAP) method were used to estimate the antioxidant activity of Jinsimyusaeng-hwan. C2C12 myoblasts were used to reevaluate the antioxidant effects. And apoptosis analysis, mitochondrial membrane potential analysis, measurement of intracellular reactive oxygen species levels were conducted to investigate antioxidant activity of Jinsimyusaeng-hwan. Results : In comparison of DPPH free radical and ABTS cationic radical scavenging activity, it increased as the concentration of water extracts of Jinsimyusaeng-hwan (WEJ) and 70% ethanol extracts of Jinsimyusaenghwan (EEJ) increased. In the results of comparing the total phenol content and reducing power using the FRAP method, extracts with high total phenol content also showed high reducing power. In comparison of the protective effect against H2 O2 -induced oxidative stress in C2C12 myoblasts, WEJ had no significant effect, but the EEJ inhibited H2 O2 -mediated cytotoxicity in a concentration-dependent manner. The cytotoxic protective effect of EEJ against oxidative stress in C2C12 myoblasts was correlated with their inhibitory effects on H2 O2 -induced apoptosis and cell cycle arrest. In H2 O2 -treated C2C12 myoblasts, the apoptosis inhibitory effects of EEJ were associated with suppression of mitochondrial dysfunction and DNA damage. The protective effect of EEJ against H2 O2 -induced oxidative stress in C2C12 myoblasts were directly related to the inhibition of ROS generation. Conclusion : Jinsimyusaeng-hwan extracts have cytotoxic protective effects against oxidative stress, and it was better in 70% ethanol extract than in water extract.

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