Moxibustion Alleviates Ferroptosis-Associated Alterations and Promotes the Recovery of Gastrointestinal Function in a Mouse Model of Postoperative Ileus

Background/Objectives: Postoperative ileus (POI) is a common complication after abdominal surgery with limited effective therapeutic options. Methods: We first performed transcriptomic screening of publicly available intestinal manipulation (IM)/POI-related datasets (GSE134942, GSE164909, and GSE263125) using signature scoring, differential expression, and pathway enrichment with cross-cohort validation. We then evaluated moxibustion at Shenque (CV8) in an IM-induced POI mouse model. Mice were randomized into Sham, POI, MOX, and POI + MOX groups; moxibustion (1.5 cm distance, 20 min/day) was applied for 4 weeks. Gastrointestinal transit was assessed by FITC-dextran, intestinal injury by H&E staining, inflammatory mediators by ELISA (COX-2, IL-1β, IL-6, TNF-α), barrier markers by ZO-1 and occludin expression (qRT-PCR, immunoblotting, immunohistochemistry), and ferroptosis-related alterations by MDA, Fe2+, GSH and GPX4. Results: Transcriptomic screening consistently highlighted inflammation activation and barrier suppression after IM and nominated ferroptosis-associated remodeling as a reproducible feature across independent mouse cohorts with directional concordance in a human surgical dataset. Moxibustion improved transit, attenuated intestinal injury, reduced COX-2 and pro-inflammatory cytokines, partially restored ZO-1 and occludin, and alleviated a pro-ferroptotic state by lowering MDA and Fe2+, increasing GSH, and upregulating GPX4. Conclusions: These results indicate that ferroptosis emerged from public-data screening and that moxibustion promotes POI recovery, at least in part, by counteracting an IM-associated inflammation-barrier impairment-ferroptosis axis.