Magnolol (Mag) is a promising natural compound with therapeutic potential for ulcerative colitis (UC). Here we designed and fabricated an oral food-grade whey protein isolate-Tremella fuciformis polysaccharides (WPI-TFPS) stabilized pickering emulsions to encapsulate Mag (Mag-WPI-TFPS) for targeted treatment of UC. With the assistance of the WPI-TFPS, pickering emulsions were well encapsulated and formed stable microparticles with a particle size of approximately 9.49 ± 0.047 μm, a 93.63 ± 0.21 % encapsulation efficiency and a loading efficiency of 21.53 ± 0.01 %. In vitro, the formulation exhibited sustained-release properties in simulated colon fluid with a cumulative release rate of 60.78 % at 48 h. In vivo, the Mag-WPI-TFPS specifically accumulated in the colon tissue for 24 h with stronger fluorescence intensity, which demonstrated that TFPS and WPI had a good adherence ability to inflamed mucosa by electrostatic attraction and ligand-receptor interactions. As expected, compared with Free-Mag, the oral administration of Mag-WPI-TFPS remarkably alleviated the symptoms of UC and protected the colon tissue in DSS-induced UC mice. More importantly, WPI-TFPS enhanced gut microbiota balance by increasing the diversity and relative abundances of Lactobacillaceae and Firmicutes. Overall, this study presents a convenient, eco-friendly, food-derived oral formulation with potential as a dietary supplement for targeted UC treatment.