This study aimed to develop low glycemic index (GI) noodles by incorporating probiotic-fermented modified wheat bran dietary fiber (DF), resistant starch (RS), and gluten (G), further investigate their collaborative effects on construction, molecular network, physicochemical, functional properties, and digestion of noodles in vitro and in vivo. Results indicated that optimized formula (10% DF/20% RS/10%G) significantly reduced GI by 44.2% (p < 0.05), from 88.14 (only wheat flour, WF control) to 49.19 through ternary cooperation mechanism: DF enhanced glucose capture, RS formed crystalline domains and repaired gluten network, while G reinforced the matrix via disulfide crosslinking. Compared to WF control, the novel formulation increased maximum tensile strength by 63.64% and reduced glucose AUC in vivo by 6.1%.Threshold analysis established compositional limits (DF ≤ 10%, G ≤ 10%) to prevent structural disruption. This work provides combined action of DF, RS, and G, suggesting the strategy for functional low - GI staple development.