Alimentary obesity is a risk factor for the development of many pathophysiological conditions in various organs, including the pancreas. Thus, the study of mechanisms, clinical symptoms and ways to prevent the development of fatty degeneration of pancreas at obesity is a current direction of research. The aim of our work was to study the influence of L-tryptophan on the morphofunctional changes of the pancreas of rats with diet-induced obesity and to evaluate the possibility of its use for the prevention of the development of the gland fatty degeneration. The study was conducted in male Wistar rats, which were 3 months old at the experiment beginning. Histologic preparations were made from pancreas tissue samples using a standard method. Morphometric measurements were performed on digital images using “Image J” software. In biochemical studies, we determined concentration of glucose in blood serum and of triglycerides, lipids and cholesterol in pancreas tissue samples. It was found that rats fed a high-fat, high-carbohydrate diet showed marked sings of developing alimentary obesity. This was evidenced by a significant increase in the weight of visceral fat (by 147%) and obesity index (by 129%). The exposure of rats to a high-calorie diet resulted in the emergence of distinct signs indicating hypofunction in both the exocrine and, to a greater extent, endocrine sections of the pancreas. The administration of L-tryptophan reduced the intensity of accumulation of visceral fat and fat in the gland itself. This was evidenced by lower concentrations of lipids (by 53%) and triglycerides (by 32%) in the pancreatic tissue compared to high-calorie diet rats. In addition, L-tryptophan prevented an excessive decrease in the function of both the exocrine and endocrine parts of the gland from the harmful effects of dietary obesity. This may be of practical interest when using tryptophan and its derivatives in the clinic to prevent a decrease in gland activity in this pathology.