c-Jun NH2-Terminal Protein Kinase Phosphorylates the Nrf2-ECH Homology 6 Domain of Nuclear Factor Erythroid 2-Related Factor 2 and Downregulates Cytoprotective Genes in Acetaminophen-Induced Liver Injury in Mice
c-Jun NH2-Terminal Protein Kinase Phosphorylates the Nrf2-ECH Homology 6 Domain of Nuclear Factor Erythroid 2-Related Factor 2 and Downregulates Cytoprotective Genes in Acetaminophen-Induced Liver Injury in Mice
复制标题
c-Jun NH2-末端激酶磷酸化核因子红细胞 2 相关因子 2 的 Nrf2-ECH 同源 6 结构域并下调对乙酰氨基酚诱导的小鼠肝损伤中的细胞保护基因
DOI：
10.1002/hep.31116
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发表时间：
2020-05-01
期刊：
影响因子：
13.5
通讯作者：
Wang, Xiu Jun
中科院分区：
文献类型：
--
作者：
Chen, Yiping;Liu, Kaihua;Wang, Xiu Jun
背景和目的对乙酰氨基酚（APAP）过量可导致严重的肝损伤和肝功能衰竭。虽然c-Jun NH 2-末端激酶（JNK）的激活被认为是APAP诱导的肝损伤的机制，但由核因子红细胞2相关因子2（Nrf 2）控制的肝防御系统在减轻APAP毒性中起着核心作用。然而，在APAP诱导的肝损伤（AILI）中，两个信号通路之间的联系仍然不清楚。方法和结果在这项研究中，我们证明了暴露于APAP后小鼠肝脏中JNK的激活与Nrf 2的磷酸化和抗氧化反应元件（ARE）驱动的基因NAD（P）H：醌脱氢酶1、谷胱甘肽S-转移酶α 3、谷胱甘肽S-转移酶M1、谷胱甘肽S-转移酶M5和醛酮还原酶1C。JNK抑制剂SP 600125或通过感染表达JNK小干扰RNA的腺病毒敲低JNK，改善APAP诱导的肝毒性，并通过稳定转录因子抑制Nrf 2磷酸化和解毒酶的下调。从机制上讲，JNK拮抗Nrf 2和ARE驱动的基因表达的Kelch样ECH相关蛋白1独立的方式。生化分析表明，磷酸化JNK（P-JNK）直接与Nrf 2的Nrf 2-ECH同源（Neh）1结构域相互作用，并磷酸化Nrf 2的Neh 6结构域中的丝氨酸-天冬氨酸-丝氨酸基序1（SDS 1）区域。结论质谱分析表明，磷酸化JNK调节Nrf 2泛素化的主要磷酸化位点是mNrf 2 SDS 1区域的丝氨酸335。这项研究表明，Nrf 2是AILI中P-JNK的靶点。我们的发现可能为AILI的治疗提供一种策略。
Background and Aims Acetaminophen (APAP) overdose induces severe liver injury and hepatic failure. While the activation of c-Jun NH2-terminal kinase (JNK) has been implicated as a mechanism in APAP-induced liver injury, the hepatic defense system controlled by nuclear factor erythroid 2-related factor 2 (Nrf2) plays a central role in the mitigation of APAP toxicity. However, the link between the two signaling pathways in APAP-induced liver injury (AILI) remains unclear.Approach and Results In this study, we demonstrated that the activation of JNK in mouse liver following exposure to APAP was correlated with the phosphorylation of Nrf2 and down-regulation of the antioxidant response element (ARE)-driven genes, NAD(P)H:quinone dehydrogenase 1, glutathione S-transferase alpha 3, glutathione S-transferase M1, glutathione S-transferase M5, and aldo-keto reductase 1C. The JNK inhibitor, SP600125, or knockdown of JNK by infection of adenovirus expressing JNK small interfering RNA, ameliorated the APAP induced liver toxicity, and inhibited the phosphorylation of Nrf2 and down-regulation of detoxifying enzymes by stabilizing the transcription factor. Mechanistically, JNK antagonized Nrf2- and ARE-driven gene expression in a Kelch-like ECH-associated protein 1-independent manner. Biochemical analysis revealed that phosphorylated JNK (P-JNK) directly interacted with the Nrf2-ECH homology (Neh) 1 domain of Nrf2 and phosphorylated the serine-aspartate-serine motif 1 (SDS1) region in the Neh6 domain of Nrf2.Conclusions Mass spectrometric analysis identified serine 335 in the SDS1 region of mNrf2 as the major phosphorylation site for modulation of Nrf2 ubiquitylation by P-JNK. This study demonstrates that Nrf2 is a target of P-JNK in AILI. Our finding may provide a strategy for the treatment of AILI.