Nuclear Nrf2 Induction by Protein Transduction Attenuates Osteoclastogenesis
Nuclear Nrf2 Induction by Protein Transduction Attenuates Osteoclastogenesis
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DOI：
10.1016/j.freeradbiomed.2014.09.006
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发表时间：
2014-12-01
影响因子：
7.4
通讯作者：
Nakamura, Yoshiki
中科院分区：
文献类型：
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作者：
Kanzaki, Hiroyuki;Shinohara, Fumiaki;Nakamura, Yoshiki
据报道，活性氧 (ROS) 在 RANKL 刺激中发挥细胞内信号分子的作用。之前我们证明，通过 Nrf2 基因转移诱导细胞保护酶表达可成功改善 RANKL 依赖性破骨细胞生成。在本研究中，我们假设通过 ETGE 肽抑制 Nrf2 泛素化和降解来激活 Nrf2，从而诱导 Nrf2 依赖性细胞保护酶表达，减弱 ROS 信号传导，从而抑制 RANKL 依赖性破骨细胞生成。将含有细胞渗透性序列（七个连续精氨酸；7R-ETGE）的 ETGE 肽应用于小鼠巨噬细胞系 RAW 264.7 细胞或原代巨噬细胞培养物。 ETGE 肽可阻止 Keap1 与 Nrf2 结合。观察到 Nrf2 核易位和 Nrf2 依赖性细胞保护酶诱导。检查了 7R-ETGE 对 RANKL 依赖性细胞内 ROS 水平诱导和骨软骨生成的影响。最后，在小鼠中研究了 7R-ETGE 对 RANICL 介导的骨破坏的保护作用。 7R-ETGE剂量依赖性地诱导核Nrf2，然后在基因和蛋白质水平上诱导细胞保护酶的表达。 7R-ETGE 通过 RANKL 刺激抑制细胞内 ROS 水平的上调，破骨细胞生成减弱。特别令人感兴趣的是局部注射 7R-ETGE 改善了 RANKL 介导的骨破坏。通过蛋白质转导局部诱导核Nrf2是牙周炎和类风湿性关节炎等骨质破坏疾病的潜在新治疗靶点。 (C) 2014 年作者。由爱思唯尔公司出版
It has been reported that reactive oxygen species (ROS) play a role as intracellular signaling molecules in RANKL stimulation. Previously we demonstrated that induction of cytoprotective enzyme expression by Nrf2-gene transfer successfully ameliorated RANKL-dependent osteoclastogenesis. In the present study, we hypothesized that Nrf2 activation by inhibiting ubiquitination and degradation of Nrf2 by ETGE-peptide would induce Nrf2-dependent cytoprotective enzyme expression, attenuate ROS signaling, and thereby inhibit RANKL-dependent osteoclastogenesis. ETGE-peptide containing a cell-permeable sequence (seven consecutive arginine; 7R-ETGE) was applied to a mouse macrophage cell-line RAW 264.7 cell or a primary macrophage culture. ETGE-peptide prevents Keap1 from binding to Nrf2. Nrf2 nuclear translocation and Nrf2-dependent cytoprotective enzyme induction was observed. The effects of 7R-ETGE on RANKL-dependent induction of intracellular ROS levels and osteodastogenesis were examined. Finally, the protective effect of 7R-ETGE on RANICL-mediated bone destruction was investigated in mice. 7R-ETGE dose-dependently induced nuclear Nrf2, followed by the induction of cytoprotective enzyme expression at both the gene and protein level. 7R-ETGE inhibited upregulation of intracellular ROS levels by RANKL stimulation, and osteoclastogenesis was attenuated. Of particular interest was that local injection of 7R-ETGE ameliorated RANKL-mediated bone destruction. Local induction of nuclear Nrf2 by protein transduction is a potential novel therapeutic target for bone destruction diseases such as periodontitis and rheumatoid arthritis. (C) 2014 The Authors. Published by Elsevier Inc.