Inhibition of Ref-1 Stimulates the Production of Reactive Oxygen Species and Induces Differentiation in Adult Cardiac Stem Cells
Inhibition of Ref-1 Stimulates the Production of Reactive Oxygen Species and Induces Differentiation in Adult Cardiac Stem Cells
复制标题
DOI：
10.1089/ars.2008.2195
复制
发表时间：
2009-03-01
影响因子：
6.6
通讯作者：
Das, Dipak K.
中科院分区：
文献类型：
--
作者：
Gurusamy, Narasimman;Mukherjee, Subhendu;Das, Dipak K.
氧化还原效应蛋白-1（Ref-1）在DNA修复和氧化还原调节多种转录因子中起重要作用。在本研究中，我们研究了Ref-1在维持氧化还原状态和存活能力的成人心脏干细胞的挑战与亚毒性水平的H2 O2下抑制Ref-1的RNA干扰的作用。用低浓度H2 O2处理心脏干细胞通过Akt磷酸化诱导Ref-1介导的存活信号。然而，Ref-1抑制后H2 O2处理通过激活NADPH氧化酶的组分（如p22（phox）、p47（phox）和Nox 4）广泛诱导细胞内活性氧（ROS）水平。心脏分化标志物（Nkx2.5、MEF 2C和GATA 4）和细胞凋亡引起的细胞死亡在Ref-1 siRNA中显著升高，随后是H2 O2处理的干细胞。此外，Ref-1的抑制增加了p53的水平，但降低了Akt的磷酸化，Akt是一种参与生存信号传导的分子。用ROS清除剂N-乙酰-L-半胱氨酸处理减弱Ref-1 siRNA介导的NADPH氧化酶活化和心脏分化。综上所述，这些结果表明Ref 1在维持心脏干细胞的氧化还原状态中起重要作用，并保护它们免受氧化损伤介导的细胞死亡和分化。抗氧化剂。氧化还原信号。11，589-599。
Redox effector protein-1 (Ref-1) plays an essential role in DNA repair and redox regulation of several transcription factors. In the present study, we examined the role of Ref-1 in maintaining the redox status and survivability of adult cardiac stem cells challenged with a subtoxic level of H2O2 under inhibition of Ref-1 by RNA interference. Treatment of cardiac stem cells with a low concentration of H2O2 induced Ref-1-mediated survival signaling through phosphorylation of Akt. However, Ref-1 inhibition followed by H2O2 treatment extensively induced the level of intracellular reactive oxygen species (ROS) through activation of the components of NADPH oxidase, like p22(phox), p47(phox), and Nox4. Cardiac differentiation markers (Nkx2.5, MEF2C, and GATA4), and cell death by apoptosis were significantly elevated in Ref-1 siRNA followed by H2O2-treated stem cells. Further, inhibition of Ref-1 increased the level of p53 but decreased the phosphorylation of Akt, a molecule involved in survival signaling. Treatment with ROS scavenger N-acetyl-L-cysteine attenuated Ref-1 siRNA-mediated activation of NADPH oxidase and cardiac differentiation. Taken together, these results indicate that Ref1 plays an important role in maintaining the redox status of cardiac stem cells and protects them from oxidative injury-mediated cell death and differentiation. Antioxid. Redox Signal. 11, 589-599.