Rationale: The surge of severe liver damage underscores the necessity for identifying new targets and therapeutic agents. Endoplasmic reticulum (ER) stress induces ferroptosis with Gα₁₂ overexpression. NF-κB essential modulator (NEMO) is a regulator of inflammation and necroptosis. Nonetheless, the regulatory basis of NEMO de novo synthesis and its impact on hepatocyte ferroptosis need to be established. This study investigated whether Nrf2 transcriptionally induces IKBKG (the NEMO gene) for ferroptosis inhibition and, if so, how NEMO induction protects hepatocytes against ER stress-induced ferroptosis./nMethods: Experiments were conducted using human liver tissues, hepatocytes, and injury models, incorporating NEMO overexpression and Gα₁₂ gene modulations. RNA sequencing, immunoblotting, immunohistochemistry, reporter assays, and mutation analyses were done./nResults: NEMO downregulation connects closely to ER and oxidative stress, worsening liver damage via hepatocyte ferroptosis. NEMO overexpression protects hepatocytes from ferroptosis by promoting glutathione peroxidase 4 (GPX4) expression. This protective role extends to oxidative and ER stress. Similar shifts occur in nuclear factor erythroid-2-related factor-2 (Nrf2) expression alongside NEMO changes. Nrf2 is newly identified as an IKBKG (NEMO gene) transactivator. Gα₁₂ changes, apart from Nrf2, impact NEMO expression, pointing to post-transcriptional control. Gα₁₂ reduction lowers miR-125a, an inhibitor of NEMO, while overexpression has the opposite effect. NEMO also counters ER stress, which triggers Gα₁₂ overexpression. Gα₁₂'s significance in NEMO-dependent hepatocyte survival is confirmed via ROCK1 inhibition, a Gα₁₂ downstream kinase, and miR-125a. The verified alterations or associations within the targeted entities are validated in human liver specimens and datasets originating from livers subjected to exposure to other injurious agents./nConclusions: Hepatic injury prompted by ER stress leads to the suppression of NEMO, thereby facilitating ferroptosis through the inhibition of GPX4. IKBKG is transactivated by Nrf2 against Gα₁₂ overexpression responsible for the increase of miR-125a, an unprecedented NEMO inhibitor, resulting in GPX4 induction. Accordingly, the induction of NEMO mitigates ferroptotic liver injury.

中文翻译：

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Nrf2 和 miR-125a 对 NEMO 的双重调节可抑制铁死亡并保护肝脏免受内质网应激诱导的损伤

理由：严重肝损伤的激增强调了确定新靶点和治疗药物的必要性。内质网 (ER) 应激可诱导铁死亡和 Gα ₁₂过度表达。 NF-κB 必需调节剂 (NEMO) 是炎症和坏死性凋亡的调节剂。尽管如此，NEMO从头合成的调控基础及其对肝细胞铁死亡的影响仍需确定。本研究调查了 Nrf2 是否通过转录诱导IKBKG（NEMO 基因）来抑制铁死亡，如果是的话，NEMO 诱导如何保护肝细胞免受 ER 应激诱导的铁死亡。/n方法：使用人类肝组织、肝细胞和损伤模型进行实验，结合 NEMO 过表达和 Gα ₁₂基因调节。进行了 RNA 测序、免疫印迹、免疫组织化学、报告基因检测和突变分析。/n结果： NEMO 下调与 ER 和氧化应激密切相关，通过肝细胞铁死亡而恶化肝损伤。 NEMO 过表达通过促进谷胱甘肽过氧化物酶 4 (GPX4) 表达来保护肝细胞免于铁死亡。这种保护作用延伸到氧化应激和内质网应激。与 NEMO 变化一起，核因子红细胞 2 相关因子 2 (Nrf2) 表达也发生类似的变化。 Nrf2 被新鉴定为IKBKG（NEMO 基因）反式激活因子。除了 Nrf2 之外，Gα _{12 的}变化也会影响 NEMO 表达，表明转录后控制。 Gα ₁₂减少会降低 NEMO 抑制剂 miR-125a，而过度表达则会产生相反的效果。 NEMO 还可以对抗 ER 应激，从而触发 Gα ₁₂过度表达。 Gα _{12在 NEMO 依赖性肝细胞存活中的重要性通过 ROCK1 抑​​制、Gα 12}下游激酶和 miR-125a得到证实。目标实体内经过验证的改变或关联在人类肝脏标本和源自暴露于其他有害物质的肝脏的数据集中得到了验证。/n结论： ER应激引起的肝损伤导致 NEMO 的抑制，从而通过抑制GPX4。IKBKG被 Nrf2 反式激活，对抗 Gα ₁₂过度表达，导致 miR-125a（一种前所未有的 NEMO 抑制剂）增加，从而诱导 GPX4。因此，NEMO 的诱导可减轻铁死亡性肝损伤。