Microplastics are difficult to degrade and widespread environmental pollutants. Coastal areas are hardest hit of microplastic pollution as they receive significant amounts of microplastics discharged from inland sources. Golden pompano (Trachinotus blochii) is a high commercial valuable marine aquaculture fish species, most of the golden pompano are raised in coastal areas, which means they are at significant risk of exposure to microplastics. Therefore, we exposed golden pompano to 10 μg/L, 100 μg/L and 1000 μg/L of 5 μm spherical polystyrene microplastics and conducted a 14-day stress experiment. Histopathology results showed the intestinal villi shrank. The 16s sequencing analysis revealed that microplastics significantly impacted the abundance and community structure of intestinal microorganisms, which may affect the metabolic function of the gastrointestinal tract. Metabolomics sequencing of the intestinal contents showed that microplastics caused disruptions in lipid, glucose, and amino acid metabolism, thus compromising the normal digestion and absorption functions in the intestinal system. In addition, the activation of various pathways, including the intestinal endocrine system, proline metabolism, and signal transduction, which can lead to the occurrence of several diseases. This study combined various methods to investigate the adverse effects of microplastics on intestinal digestion and absorption, and provided new insights into the toxic mechanisms of microplastics.