Ren-Na Wu, Qinglin Bao, Yasula Ba, Chaoketu Saiyin, Lengge Si, A Runa
Objective: This study evaluated the antidepressant-like effects of Mongolian medical warm acupuncture (MMWA) in a chronic unpredictable mild stress (CUMS) model and examined its mechanistic involvement in the gut microbiota-metabolite-barrier axis, representing a novel multi-omics investigation of this traditional therapy. Methods: Control, CUMS, and MMWA rats (n = 9/group) were assessed using sucrose preference, open-field activity, and Morris water maze tasks. Gut microbiota, fecal metabolites, and intestinal barrier markers were measured by 16S rRNA sequencing, UPLC-MS/MS metabolomics, qRT-PCR, and Western blotting. Results: MMWA improved CUMS-induced deficits, increasing sucrose preference (p < 0.01), enhancing locomotor activity (p < 0.01), and reducing escape latency (p < 0.05). Treatment restored microbial diversity and increased beneficial short-chain fatty acid (SCFA)-producing genera, including Lactobacillus and Prevotella (p < 0.05). Metabolomic analysis showed recovery of key neuroactive metabolites such as taurine and arginine (adjusted p < 0.05). MMWA also enhanced intestinal barrier integrity by upregulating Occludin, TJP1/ZO-1, and Claudin-4 (p < 0.001). Associations across microbiota-metabolite pathways reflected coordinated restoration. Conclusion: MMWA alleviates depressive-like behaviors by reshaping gut microbiota, normalizing metabolic profiles, and strengthening the intestinal barrier. These findings support its potential as a complementary approach for depressive-like conditions and highlight a mechanistic link involving the microbiota-metabolite-barrier axis.