In this study, the hypoglycemic effects and mechanisms of different intervention substances (Trichosanthis Radix resistant starch, rutin, Trichosanthis Radix resistant starch-rutin complex) were investigated on C57BL/6 diabetic mice induced by high glucose-fat diet combined with streptozotocin. The results indicate that the Trichosanthis Radix resistant starch (T-RS) group showed superior effect compared to the rutin group in terms of increasing body weight, lowering blood glucose, regulating oral glucose tolerance, reducing insulin levels, promoting glycogen and lipid metabolism; while rutin group exhibited a better inhibitory effect on oxidative stress related to diabetes. Compared to the rutin group and T-RS group, the Trichosanthis Radix resistant starch -rutin complex (RuT-RS) group showed more significant regulatory effects on liver glycogen, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and glutathione peroxidase (P < 0.05). In summary, the combination of both substances can effectively improve the capacity for glucose and lipid metabolism regulation and antioxidant ability in C57BL/6 diabetic mice. Our research results show that RuT-RS has potential applications in improving type 2 diabetes and broadens the application scope of T-RS across various sectors, including food, dietary supplements, and health products.