机构：[1]Department of Neurosurgery, Yijishan Hospital, Wannan Medical College, Wuhu, China.[2]Department of Neurosurgery, The First People’s Hospital of Kunshan, Jiangsu University, Suzhou, China.[3]Department of Endocrinology, The Second Affiliated Hospital of Soochow University, Suzhou, China.[4]Department of Neurosurgery, Kunshan Hospital of Traditional Chinese Medicine, Kunshan Affiliated Hospital, Suzhou, China.
Background: Four-octyl itaconate (OI), the itaconate's cell-permeable derivative, can activate Nrf2 signaling via alkylation of Keap1 at its cysteine residues. The current study tested the potential neuroprotective function of OI in hydrogen peroxide (H2O2)-treated neuronal cells. Methods: SH-SY5Y neuronal cells and epigenetically de-repressed (by TSA treatment) primary murine neurons were treated with OI and/or H2O2. Nrf2 pathway genes were examined by Western blotting assay and real-time quantitative PCR analysis. Neuronal cell death was tested by the LDH and trypan blue staining assays. Apoptosis was tested by TUNEL and Annexin V assays. Results: In SH-SY5Y neuronal cells and primary murine neurons, OI activated Nrf2 signaling, causing Keap1-Nrf2 disassociation, Nrf2 protein stabilization and nuclear translocation, as well as expression of Nrf2-regulated genes (HO1, NQO1 and GCLC) and ninjurin2 (Ninj2). Functional studies showed that OI attenuated H2O2-induced reactive oxygen species (ROS) production, lipid peroxidation and DNA damage as well as neuronal cell death and apoptosis. shRNA-mediated knockdown, or CRISPR/Cas9-induced knockout of Nrf2 almost abolished OI-induced neuroprotection against H2O2. Keap1 is the primary target of OI. Keap1 knockout by CRISPR/Cas9 method mimicked and abolished OI-induced actions in SH-SY5Y cells. Introduction of a Cys151S mutant Keap1 in SH-SY5Y cells reversed OI-induced Nrf2 activation and anti-H2O2 neuroprotection. Conclusions: OI activates Keap1-Nrf2 signaling to protect SH-SY5Y cells and epigenetically de-repressed primary neurons from H2O2 in vitro.
基金：
This work is supported by the Clinical and basic researches of brain disease
(KYC004), the Science Foundation of Wannan Medical College (WK2017F04)
and Kunshan Science Project (KS1644).
第一作者机构：[2]Department of Neurosurgery, The First People’s Hospital of Kunshan, Jiangsu University, Suzhou, China.
共同第一作者：
通讯作者：
通讯机构：[1]Department of Neurosurgery, Yijishan Hospital, Wannan Medical College, Wuhu, China.
推荐引用方式(GB/T 7714)：
Hua Liu,Yu Feng,Min Xu,et al.Four-octyl itaconate activates Keap1-Nrf2 signaling to protect neuronal cells from hydrogen peroxide[J].CELL COMMUNICATION AND SIGNALING.2018,16(1):81.doi:10.1186/s12964-018-0294-2.
APA：
Hua Liu,Yu Feng,Min Xu,Jian Yang,Zhichun Wang&Guangfu Di.(2018).Four-octyl itaconate activates Keap1-Nrf2 signaling to protect neuronal cells from hydrogen peroxide.CELL COMMUNICATION AND SIGNALING,16,(1)
MLA：
Hua Liu,et al."Four-octyl itaconate activates Keap1-Nrf2 signaling to protect neuronal cells from hydrogen peroxide".CELL COMMUNICATION AND SIGNALING 16..1(2018):81
