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Rutin and rutin-rich buckwheat are commonly used as alternative medicines due to their wide range of health benefits. The present study aims to investigate their anti-diabetes effects and the association with glucagon-like peptide 1 (GLP-1) levels. A murine model of type 2 diabetes (T2D), induced by a high-fat diet combined with streptozocin injection, was utilized. T2D mice were fed 3% buckwheat, 10% buckwheat, or 0.1% rutin as part of their diet for 4 weeks. Dipeptidyl peptidase-4 assays, secretin tumor cell line (STC-1) cells, and GLP-1 receptor-knockout mice were employed for mechanistic studies. Dietary consumption of 10% buckwheat and 0.1% rutin provided beneficial outcomes including lowering blood glucose levels and HbA1c, enhancing HOMA-B%, improving glucose tolerance and pyruvate tolerance. Serum GLP-1, ghrelin, leptin, and adiponectin levels were also increased at the end of the treatment. Increased GLP-1 levels and the improvement of β-cell function were closely associated with the blood glucose-lowering effects observed. Hypoglycemic effects of buckwheat and rutin supplement were abolished in diabetic GLP-1 receptor knockout mice. In conclusion, rutin and related herbal materials are expected to provide health benefits due to their ability to modulate endogenous GLP-1 levels in T2D mice.