Electroacupuncture Alleviates Oxidative Stress and Cuproptosis in Metabolic Dysfunction-Associated Steatotic Liver Disease Mice Through Activation of the NRF2/ARE Pathway

Junyuan Deng,1 Cai Liao,1 Hejing Liu,1 Xin Tang,1 Yan Yang,1 Yunhao Yang,2 Xiao Guo,1 Shanshan Zhang,1 Kehan Xing,1 Mei Liao,1 Chenglin Tang1 1College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing, People’s Republic of China; 2College of Acupuncture and Tuina, Chongqing University of Chinese Medicine, Chongqing, People’s Republic of ChinaCorrespondence: Chenglin Tang, Email [email protected]: Oxidative stress and the newly characterized mode of regulated cell death, cuproptosis, drive the progression of metabolic dysfunction-associated steatotic liver disease (MASLD). Electroacupuncture (EA) is a promising non-drug treatment, but its mechanisms are unclear. This study investigated the benefits of EA in a high-fat diet (HFD)-induced MASLD mouse model, focusing on the NRF2/ARE pathway, cuproptosis, and the gut–liver axis.Methods: MASLD was established in C57BL/6J mice via a 16-week HFD. Mice were arbitrarily categorized into four groups: control (C, normal chow diet), model (M, HFD), EA (HFD + EA), and EA plus NRF2 inhibitor (EM, HFD + EA + inhibitor). A 4-week intervention was then conducted. Post-intervention, body weight alterations were tracked, and serum, liver tissue, and fecal specimens were gathered for subsequent examination.Results: EA treatment significantly improved the metabolic profile, reducing body weight, lipids, and hepatic inflammation. It attenuated oxidative stress by enhancing antioxidant capacity and activated the NRF2/ARE pathway. EA also modulated cuproptosis-related genes (upregulating dihydrolipoamide S-acetyltransferase (Dlat), downregulating solute carrier family 31 member 1 (Slc31a1), ferredoxin reductase (Fdx1), and heat shock protein 70 (Hsp70)). Furthermore, EA remodeled the gut microbiota (increasing Limosilactobacillusand Ligilactobacillus) and elevated related metabolites.Conclusion: EA ameliorates MASLD through multiple mechanisms, including regulating lipid metabolism, attenuating inflammation and oxidative stress, activating NRF2, suppressing cuproptosis, and altering the gut–liver axis, collectively contributing to hepatic improvement. Keywords: gut microbiota, metabolomics, hepatic lipid metabolism, inflammation, mitochondrial dysfunction, antioxidant therapy