Glucagon-like peptide-1 (GLP-1), one of the expression products of the proglucagon (pg) gene, is an incretin mainly secreted by the gastrointestinal system. In mammals, GLP-1 has hypoglycemic and food-inhibiting effects; while in some fish species, it has been confirmed to increase blood glucose by promoting gluconeogenesis and stimulating glycogenolysis. In order to more deeply understand the role of GLP-1 in the process of glycometabolism in herbivorous fish, the pg gene was cloned from Spinibarbus denticulatus to obtain its sequence characteristics, and the changes in blood glucose level and pg gene expression in S. denticulatus were further explored by feeding with three kinds of carbohydrates and intraperitoneal injection of GLP-1. Basal and temporal blood glucose levels and pg gene expression of S. denticulatus (91.68 ± 10.79 g) were measured at 0, 1, 3, 5, 7, and 12 h after oral administration ( n = 4). Then, the changes of blood glucose levels and pg and glucokinase (gk) gene expressions of S. denticulatus (94.29 ± 10.82 g) were determined at 0, 30, 60, and 120 min after intraperitoneal injection ( n = 4). It was shown that polysaccharides could induce the upregulation of pg gene expression faster than monosaccharides and stimulate the secretion of GLP-1 in the intestine. Intraperitoneal injection of GLP-1 peptide rapidly raised blood glucose levels, and pg gene expression in the anterior intestine, whole brain, and hepatopancreas decreased continuously after 30 minutes. These results showed that S. denticulatus might inhibit the excessive accumulation of blood glucose by reducing the expression of the pg gene and increasing the expression of gk gene in a short time. It was speculated that GLP-1 of S. denticulatus might have a “gut-brain-liver” pathway similar to mammals in glycemia regulation. Therefore, this study provided a novel perspective for explaining the functional differences of GLP-1 in herbivorous fish and mammals.