In summary, we found that methane-rich saline had a protective effect on acetic acid-induced ulcerative colitis, relying on the ability to alleviate oxidative stress and inflammation by inhibiting the TLR4/NF-κB/MAPK signaling pathway and promoting IL-10/JAK1/STAT3-mediated anti-inflammatory response. The gene discussed is TLR4; the disease is ulcerative colitis.