This study investigated the in vitro gastrointestinal protective effects of a multi-component food bioactive formulation composed of turmeric, manuka honey, sea buckthorn, black pepper, and chocolate chili extracts. The formulation was evaluated in peripheral blood mononuclear cells, gastric epithelial cells, and esophageal epithelial cells for its influence on lipopolysaccharide-induced cytokine secretion, mucin (MUC5B) production, and tight-junction protein (ZO-1) expression, as well as its antimicrobial activity against Helicobacter pylori. Compared with single ingredients or partial combinations, the complete formulation demonstrated synergistic activity by attenuating proinflammatory cytokine release, reinforcing epithelial barrier integrity, normalizing mucin secretion, and inhibiting H. pylori growth. These findings provide mechanistic evidence that multi-component food bioactive formulations can contribute to gastrointestinal protection through complementary anti-inflammatory and antimicrobial mechanisms, supporting their potential as functional strategies for maintaining gut health.