Kolaviron (KV) as an established hypoglycemic agent in both normal and experimental diabetic rat among other effects was investigated for its mechanism of hypoglycemia on both normal and diabetic Wister rats with emphasis on the role of ileum in glucose uptake. Female rats weighing between 150 -180g were used and grouped into 4 of six animals each. Group 1 and 2 represents the control (non-diabetic) and untreated diabetic respectively while groups 3 and 4 were treated with 200mg/kg KV and 5mg/kg Glibenclamide (GB). The study involved three phases, diabetes was induced by single intraperitoneal injection of 120mg/kg b. wgt. alloxan and animals with fasting blood glucose greater than 200mg/dl were considered for the study. Prior to each phase, rats were fasted for 12 hour with free access to water. Phase 1 was conducted to check the effect of various treatments on blood glucose of fasted control and diabetic rats while Phases 2 and 3 were to investigate the route of glucose utilization following various treatments via the arterial -venous (A-V) differences and ileum glucose uptake respectively. Rats were anaesthetized with 1.9% ether on a cotton ball inside a conical tube without direct contact with the animals prior to laparotomy in Phases 2 and 3 following which 4ml of modified Krebs solution was infused between 2 ligated ends of ileum(15-20cm). Drops of blood/ luminal samples were taken from the lateral tail vein, abdominal aorta and mesenteric veins and the isolated ileum lumen for phases 1, 2 and 3 respectively. Analysis was done using standardized digital glucometer analyzer. Subsequently, animals were sacrificed with excess ether in a closed chamber. Data were analyzed using students ‘t’ test and ANOVA, results were significant at P<0.05. Phase 1 showed significant decrease (P<0.05, n=6) in blood glucose concentration of KV treated group(218±9.2mg/dl) and GB treated group (311±8.1 mg/dl) compared to the Control (81±3.1 mg/dl) and Diabetic (non-treated) (411±14 mg/dl) after 2 hours of substances administration. The A-V study recorded a 14.1%, 27.6% and 19.3% decrease in A-V difference of blood glucose concentration in the untreated diabetics, KV treated and GB treated groups respectively compared to the control after 2 hours of treatments. The ileum glucose absorption increased significantly (P<0.05, n=6) in the KV treated (12.67±1.2mg/dl/cm, 18.12±1.6mg/dl/cm) and GB treated (10.41±0.8 mg/dl/cm, 16.51±1.1 mg/dl/cm) when compared with the Control (8.66±0.9 mg/dl/cm, 11.61±1.6 mg/dl/cm) and untreated diabetic (9.91±0.8 mg/dl/cm, 11.13±0.9 mg/dl/cm) after 90 and 120 minutes treatment respectively. We concluded that KV could exert its profound hypoglycemic effect through other routes other than stimulation of the pancreatic β-cells as buttressed by increase ileum glucose absorption reported.