Functional foods enriched with plant-derived bioactive compounds have emerged as promising complementary dietary strategies to improve glycemic regulation. The aim of this study was to evaluate the physicochemical properties, sensory acceptance, and microbiological characteristics of a functional yogurt enriched with Moringa oleifera leaf extract, as well as its effect on blood glucose levels in an alloxan-induced diabetic rat model. Phytochemical characterization of the hydroethanolic leaf extract by HPLC and direct-infusion ESI-IT-MS² revealed phenolic compounds, including chlorogenic acid, rutin, quercetin, and kaempferol, together with additional glycosylated flavonoids. M. oleifera extract increased the yogurt’s protein content. Sensory evaluation indicated moderate acceptance compared to natural yogurt, with color being the least-accepted attribute. Microbiological analysis showed no inhibitory effect against representative bacterial genera of the gastrointestinal microbiota under the experimental conditions evaluated. In vivo evaluation in alloxan-induced diabetic female Wistar rats demonstrated a significant reduction in blood glucose levels, from 396.50 ± 17.85 mg/dL to 254.50 ± 32.79 mg/dL after 15 days of treatment. These findings support further investigation of M. oleifera functional yogurt as a potential functional food in the context of glycemic control.