Mus musculus
Objective: This study explores the therapeutic effects of Artesunate (ART) on Sjgren's syndrome in NOD/Ltj mice by modulating ferroptosis in salivary gland epithelial cells (SGECs). Methods: Female NOD/Ltj mice at 8 weeks of age were randomly divided into two groups: a model group treated with saline and an ART group receiving oral gavage of ART. Daily water intake, weekly salivary flow rate, and body weight were monitored. At the end of the 8-week treatment period, mice were euthanized, and spleen and submandibular gland indices were assessed. scRNA-seq was used to analyze the characteristic profiles of SS patients, while RNA-seq evaluated inflammatory pathway signaling responses following ART treatment. Histological examination included HE staining of submandibular glands, scoring based on lymphocytic infiltration. Western blotting detected KEAP1, TFRC, xCT, NRF2, and GPX4 expression levels in submandibular glands and SGECs. Immunofluorescence was employed to examine IgG, C3, KEAP1, TFRC, NRF2, xCT, and GPX4 expression in submandibular glands, as well as ROS and JC-1 RED/GREEN levels in SGECs. Molecular docking assessed the affinity between ART and KEAP1, and transmission electron microscopy evaluated mitochondrial morphology. Results: scRNA-seq and systems biology analyses revealed robust activation of ferroptosis signaling pathways post-ART treatment. In vitro and in vivo studies further confirmed that ART inhibited the ubiquitination and degradation of NRF2 by KEAP1, upregulated xCT and GPX4 protein expression, and suppressed TFRC levels. Consequently, ART significantly protected SGECs from ferroptosis in NOD/Ltj mice, effectively preventing glandular damage and maintaining functional integrity.