OBJECTIVE: Diabetes mellitus (DM) is a chronic metabolic disorder characterized by elevated blood glucose levels due to insulin deficiency or resistance. While conventional treatments are effective, integrating nutraceuticals that aid in blood sugar control is crucial for patients with low compliance or prediabetes. This study aimed to assess the efficacy of kefir-derived exopolysaccharides (KEPS) in ameliorating type 2 DM in rats induced by streptozotocin (STZ) and a high-fat diet. RESULTS: Eight-week-old male SD rats were fed a high-fat diet for 4 weeks and subsequently administered STZ (35 mg/kg body weight) via intraperitoneal injection to induce type 2 diabetes. Diabetic rats were randomly assigned to three groups: untreated diabetes (mock), diabetic rats + high-dose KEPS (STZ-KEPSH), and diabetic rats + low-dose KEPS (STZ-KEPSL), and treated for an additional 4 weeks. An age-matched SD rat group without STZ induction was used as a control. Our findings demonstrate that high-dose KEPS administration in diabetic rats enhances hepatic glucose uptake and stabilizes blood glucose levels by upregulating GLUT2 protein expression and PI3k phosphorylation in liver cells. KEPS mitigates lipid production, reduces β cell damage, and preserves islet cell integrity, thereby improving insulin sensitivity and alleviating diabetes symptoms. High-dose KEPS treatment also exhibits less renal enlargement and lower kidney-to-body weight ratio than the diabetic group. CONCLUSION: Exopolysaccharides derived from kefir show promising potential in managing type 2 diabetes mellitus in rats, suggesting a viable nutraceutical therapy. Further research is needed to validate these findings in human subjects.