Ulcerative colitis is a chronic inflammatory condition of the intestine characterized by mucosal damage and a compromised epithelial barrier. This study explored the protective and therapeutic potential of laminaran derived from the brown seaweed Undaria pinnatifida in promoting mucin secretion and restoring mucosal barrier integrity. Physicochemical analysis revealed laminaran as having a beta-(1 --> 3)-linked glucose backbone with beta-(1 --> 6)-linked branches and a molecular weight of 14.41 kDa. In vitro experiments revealed that laminaran enhanced the expression of mucin-related proteins in a lipopolysaccharide-induced LS174T model. Laminaran also upregulated the expression of sulfotransferases, which are essential for mucin sulfation, and promoted vesicular transport by increasing the expression of vesicle-associated membrane protein 8 and synaptosome-associated protein-23, facilitating mucin secretion. These effects are mediated through the protein kinase C (PKC) pathway, which involves PKCalpha and PKCbetaII. In an in vivo model, laminaran alleviated dextran sulfate sodium-induced colitis, increasing mucus thickness and overall intestinal barrier function. These results suggest that laminaran is a promising therapeutic agent for treating ulcerative colitis, suggesting a novel approach to restoring the mucosal barrier and reducing intestinal inflammation. This study lays the groundwork for developing laminaran-based treatments for ulcerative colitis and other intestinal diseases associated with epithelial barrier dysfunction.