Xia Xuyun, Zhang Qin, Fang Xia, Li Ling, Yang Gangyi, Xu Xiaohui, Yang Mengliu
Department of Endocrinology, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
The Key Laboratory of Laboratory Medical Diagnostics in the Ministry of Education and Department of Clinical Biochemistry, College of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Clin Transl Med. 2025 Mar;15(3):e70233. doi: 10.1002/ctm2.70233.
The epidemic of metabolic dysfunction-associated fatty liver disease linked to excessive high-fat diet (HFD) consumption has sparked widespread public concern. Nuclear factor erythroid 2-related factor 2 (NRF2) has been reported to improve glucose/lipid metabolism, liver lipid degeneration and alleviate HFD-induced inflammation. However, its pathways and mechanisms of action are not fully understood.
To confirm the effect of NRF2 on glucose/lipid metabolism in the liver, Nrf2-/- mice as well as liver-specific Nrf2 knockout mice, and AAV-TBG-Nrf2 were employed. The hyperinsulinemic-euglycemic clamp was utilized to determine the effect of NRF2 on glucose metabolism. To elucidate the effect of NRF2 on pyroptosis, we performed western blots, immunofluorescence, quantitative real-time PCR, and Flow cytometry experiments. Finally, chromatin immunoprecipitation-seq and dual-luciferase reporter assay was used to underscore the transcriptional regulatory effect of NRF2 on Gsdmd.
We found that overexpression of Nrf2 inhibited the expression of inflammatory cytokines and pyroptosis markers, including cle-Caspase1, NLRP3 and the N-terminus of gasdermin D (N-GSDMD) both in vivo and in vitro, while Nrf2 deficiency was the opposite. Specifically, with NRF2 expression up-regulated, GSDMD expression decreased and Gsdmd overexpression partially reversed the effect of Nrf2 overexpression on pro-inflammatory phenotype. Mechanistically, we demonstrate that NRF2 binds to the Gsdmd promoter at the -2110 - 1130 bp site, inhibiting the GSDMD expression and thereby improving glucose/lipid metabolism and liver steatosis.
Our data indicate that NRF2 is an effective inhibitor of pyroptosis and has a multi-target effect in the treatment of obesity-related metabolic diseases.
MAFLD is associated with increased hepatocytes NRF2 expression. NRF2 alleviates MAFLD by suppressing pyroptosis. NRF2 directly inhibits GSDMD expression to regulate pyroptosis. Targeting the NRF2-pyroptosis (GSDMD) axis offers a potential therapeutic strategy for MAFLD.
与过量食用高脂饮食（HFD）相关的代谢功能障碍相关脂肪性肝病的流行引发了广泛的公众关注。据报道，核因子红细胞2相关因子2（NRF2）可改善糖/脂代谢、肝脏脂质变性并减轻HFD诱导的炎症。然而，其作用途径和机制尚未完全明确。
为了证实NRF2对肝脏糖/脂代谢的影响，采用了Nrf2-/-小鼠、肝脏特异性Nrf2基因敲除小鼠以及腺相关病毒-甲状腺激素结合球蛋白-Nrf2（AAV-TBG-Nrf2）。采用高胰岛素-正常血糖钳夹技术来确定NRF2对糖代谢的影响。为了阐明NRF2对细胞焦亡的影响，我们进行了蛋白质免疫印迹、免疫荧光、定量实时聚合酶链反应和流式细胞术实验。最后，使用染色质免疫沉淀测序和双荧光素酶报告基因检测来强调NRF2对gasdermin D（Gsdmd）的转录调控作用。
我们发现，在体内和体外，Nrf2的过表达均抑制了炎性细胞因子和细胞焦亡标志物的表达，包括半胱天冬酶-1（cle-Caspase1）、NOD样受体蛋白3（NLRP3）和gasdermin D的N端（N-GSDMD），而Nrf2缺乏则产生相反的结果。具体而言，随着NRF2表达上调，GSDMD表达下降，且Gsdmd过表达部分逆转了Nrf2过表达对促炎表型的影响。机制上，我们证明NRF2在-2110至-1130 bp位点与Gsdmd启动子结合，抑制GSDMD表达，从而改善糖/脂代谢和肝脂肪变性。
我们的数据表明，NRF2是细胞焦亡的有效抑制剂，在治疗肥胖相关代谢疾病方面具有多靶点作用。
代谢功能障碍相关脂肪性肝病与肝细胞NRF2表达增加有关。NRF2通过抑制细胞焦亡减轻代谢功能障碍相关脂肪性肝病。NRF2直接抑制GSDMD表达以调节细胞焦亡。靶向NRF2-细胞焦亡（GSDMD）轴为代谢功能障碍相关脂肪性肝病提供了一种潜在的治疗策略。
