Gut hormones play a key role in the regulation of food intake, energy expenditure, glucose homeostasis, lipid metabolism, and a wide range of metabolic functions in response to food ingestion. These hormones are altered in metabolic diseases, such as obesity and type 2 diabetes, and are thus proposed to be possible targets for the prevention or treatment of these diseases. It is clear that food composition, macronutrients, and other non-nutrient components as well as the physical properties of food not only modulate the secretion of gut peptides but also modulate transcription and enteroendocrine cell differentiation, which ultimately modifies gut hormone response. The specific mechanisms or sensing machinery that respond to the different components of the diet have been studied for many years; however, over the last few years, new molecular genetic techniques have led to important advances, thereby allowing a deeper understanding of these mechanisms. This review addresses the current knowledge regarding enteroendocrine cells and how diet interacts with this machinery to stimulate and regulate the secretion of gut peptides. The potential for diet interventions as a promising strategy for modulating gut hormone responses to food ingestion and, ultimately, preventing or treating metabolic diseases is being emphasized considering that these diseases are currently a public health burden. Adv. Nutr. 3: 8-20, 2012.