| Summary: | 碩士 === 中國醫藥大學 === 藥物化學研究所碩士班 === 102 === Nrf-2 protein is a transcription factor that binds to Antioxidant-response element (ARE) regions and drives various detoxification and phase 2 metabolic enzymes, such as HO-1 and glutathione synthase. Activation of Nrf-2/ARE pathway is considered to protect cells against exogenic insults that cause diseases like diabetes and cancers. Therefore, development of Nrf-2 activators can be applied for disease prevention. In this study, NPC-TW01 and HSC-3 cells were stably transfected with ARE-luciferase reporter genes, named as TW01-9ARE and HSC3-9ARE cells, and were thus employed for screening. A panel of naphol- containing oxime derivatives was subjected to determine the Nrf-2 activation ability in the TW01-9ARE and HSC3-9ARE cells. The cytotoxicity of each compound was also measured and none of tested compounds displayed obvious cytotoxicity. Among these compounds, Naph-O204 showed the most promising effect on the activation of Nrf-2/ARE pathway and thus it was chosen for further study. The mRNA level of detoxification enzymes, HO-1, was increased, but no change of Nrf-2 and AKR1C1 mRNA was found. Furthermore, Naph-O204 significantly reduced the H2O2-induced oxidative stress in TW01-9ARE cells. In addition, the phosphorylation of Erk1/2 and Akt, which affect the Nrf-2 function, were elevated in Naph-O204-treated cells. Although mRNA and protein level of Nrf-2 were not altered after Naph-O204 treatment, the Nrf-2 phosphorylation was augmented. Moreover, blockage of PKC and PI3K-Akt pathway abolished the Naph-O204 induced Nrf-2 activation. In summary, our study demonstrated that Naph-O204 treatment can exert the cytoprotective property through stimulating PKC or Akt pathway, activating Nrf-2 protein and producing detoxification enzymes, which can be served as a drug lead for disease prevention.
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