Akkermansia Muciniphila Attenuates TNF-α-Induced Injury in Human Primary Colonic Epithelial Cells with Ferroptosis-Associated Redox Remodeling

Akkermansia muciniphila (AKK) is linked to intestinal barrier homeostasis, but whether live AKK can directly protect human colonic epithelium during a defined cytokine insult remains less clear. We tested this question in human primary colonic epithelial cells (HCoEpiC). Cells were pretreated with live AKK at different multiplicities of infection (MOIs) before tumor necrosis factor-α (TNF-α) exposure; erastin (1.76 µM) was used to increase ferroptosis pressure. Cell viability, apoptosis, interleukin-6 (IL-6)/interleukin-1β (IL-1β) concentrations, and selected ferroptosis-associated redox and epithelial-state proteins were measured by the Cell Counting Kit-8 (CCK-8) assay, flow cytometry, enzyme-linked immunosorbent assay (ELISA), and Western blotting. TNF-α reduced viability, increased apoptosis, and raised IL-6 and IL-1β concentrations. AKK pretreatment mitigated these changes, with the clearest effect at MOI 100. AKK also increased glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11), reduced acyl-CoA synthetase long-chain family member 4 (ACSL4) and NAD(P)H quinone dehydrogenase 1 (NQO1), and increased hes family bHLH transcription factor 1 (HES1), homeobox A13 (HOXA13), and homeobox B13 (HOXB13). Erastin partly weakened the AKK-associated improvements in viability, apoptosis, cytokine levels, and protein expression. These findings show that live AKK attenuates TNF-α-induced injury in primary human colonic epithelial cells. The protective pattern is accompanied by ferroptosis-associated redox remodeling and recovery of HES/HOX-associated epithelial-state markers, supporting further testing in ulcerative colitis (UC)-relevant epithelial injury models.