Dietary flavonoid genistein induces Nrf2 and phase II detoxification gene expression via ERKs and PKC pathways and protects against oxidative stress in Caco-2 cells.
Dietary flavonoid genistein induces Nrf2 and phase II detoxification gene expression via ERKs and PKC pathways and protects against oxidative stress in Caco-2 cells.
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DOI：
10.1002/mnfr.201200536
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发表时间：
2013-02
影响因子：
5.2
通讯作者：
Xiaohan Zhai;Musen Lin;Feng Zhang;Yan Hu;Xiao-mei Xu;Yubing Li;Kexin Liu;Xiaochi Ma;X. Tian
中科院分区：
文献类型：
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作者：
Xiaohan Zhai;Musen Lin;Feng Zhang;Yan Hu;Xiao-mei Xu;Yubing Li;Kexin Liu;Xiaochi Ma;X. Tian
范围黄酮类化合物具有众所周知的抗氧化、抗炎和抗癌活性。异黄酮染料木黄酮被认为是一种有效的抗氧化剂对氧化应激。虽然已经提出了多种机制，但染料木黄酮的抗氧化机制仍有待进一步研究。方法和结果在这项研究中，我们专注于Nrf 2和II相酶途径组分的表达的协同作用。通过瞬时转染、RT-PCR和免疫印迹分析研究其作用的分子机制。在Caco-2细胞中，染料木黄酮处理显著减弱了H（2）O（2）诱导的过氧化物形成;这种改善被丁噻呋砜亚胺（GCLC抑制剂）和锌原卟啉（HO-1抑制剂）逆转。金雀异黄素增加HO-1和GCLC的mRNA和蛋白表达。Genistein处理激活ERK 1/2和PKC信号通路，因此增加Nrf 2 mRNA和蛋白表达。使用PD 98059（ERK 1/2抑制剂）和GF 109203 X（PKC抑制剂）和针对Nrf 2的RNA干扰来确定ERK 1/2和PKC信号通路的作用。两种抑制剂和siNrf 2都能抑制染料木素诱导的HO-1和GCLC蛋白表达。这些结果表明ERK 1/2、PKC和Nrf 2参与了染料木素诱导HO-1和GCLC的过程。结论金雀异黄素通过EKR 1/2和PKC /Nrf 2途径上调氧化应激时HO-1和GCLC的表达。
SCOPE Flavonoids have well-known antioxidant, anti-inflammatory, and anti-cancer activities. Isoflavone genistein is considered a potent antioxidant agent against oxidative stress. Although several mechanisms have been proposed, a clear antioxidant mechanism of genistein is still remained to be answered. METHODS AND RESULTS In this study, we focused on the concerted effects on expression of Nrf2 and phase II enzyme pathway components. Transient transfection assays, RT-PCR and immunoblot analysis were performed to study its molecular mechanisms of action. In Caco-2 cells, treatment with genistein markedly attenuated H(2)O(2) -induced peroxide formation; this amelioration was reversed by buthionine sulfoximine(GCLC inhibitor) and zinc protoporphyrin(HO-1 inhibitor). Genistein increased HO-1 and GCLC mRNA and protein expression. Genistein treatment activated the ERK1/2 and PKC signaling pathway; therefore increased Nrf2 mRNA and protein expression. The roles of the ERK1/2 and PKC signaling pathway were determined using PD98059 (ERK1/2 inhibitor) and GF109203X (PKC inhibitor) and RNA interference directed against Nrf2. Both inhibitors and siNrf2 abolished genistein-induced HO-1 and GCLC protein expression. These results suggest the involvement of ERK1/2, PKC, and Nrf2 in inducing HO-1 and GCLC by genistein. CONCLUSION Our studies show that genistein up-regulated HO-1 and GCLC expression through the EKR1/2 and PKC /Nrf2 pathways during oxidative stress.