Thyroid hormone has been shown to alter glucose metabolism, however, there seems to be controversial reports on the effect of thyroxine on intestinal glucose transport. This study was conducted to investigate the effect of thyroidectomy and thyroxine on glucose transport in the small intestine. Forty rats were randomly selected into four groups of ten rats each. Groups one and two rats were thyroidectomised to make them hypothyroid after which group two rats were given thyroxine replacement of 10ug/100g b/w for thirty five days to make them euthyroid. Rats in groups three and four were sham operated thereafter group three rats were given 10ug/100g b/w thyroxine for thirty five days to make them hyperthyroid. Group four served as the control. 10mg/kg b/w Ketamine was administered intraperitoneally as anesthesia for the surgeries. On the thirty-fifth day post surgery all the animals were killed via cervical dislocation and their small intestines were harvested. 10cm length of jejunum and ileum respectively were used to make everted sacs for the invitro study. Mucosa glucose transfer (MGT), Final glucose concentration gradient (FCG) and Gut glucose uptake (GGU) were significantly higher (P< 0.05) in the hyperthyroid group and lower (P< 0.05) in the hypothyroid group compared with the control with transport in the jejunum greater (P< 0.05) than the ileum in all groups. Serosal glucose transfer (SGT) was Negative in the hyperthyroid group. SGT was significantly greater (P< 0.05) in the jejunum than the ileum. These findings suggest that thyroidectomy causes reduced glucose transport while thyroxine increases glucose transport in different segments of the small intestine with the transport in the jejunum greater than that of the ileum. The mechanisms by which these occur may be by altering the morphology of the intestinal wall, villi, as well as other absorption variables in the small intestine. Key words: Glucose transport, Thyroxine, thyroidectomy, Everted sac