Background Past research found intermittent fasting (IF) might have a protective potential in neurodegenerative diseases. So, the current work compares the impact of IF, vitamin B12, and folic acid on memory impairment induced by type 2 diabetes mellitus (T2DM). Materials and Methods Forty male albino rats were included in this study. They were distributed into 5 groups (each of 8 rats). Group I (the control group) consisted of healthy rats with normal food habits. Group II consisted of the streptozotocin-induced T2DM rats. Group III consisted of diabetic rats with IF. Group IV consisted of diabetic rats supplemented with vitamin B12 and folic acid. Group V consisted of diabetic rats with IF supplemented with vitamin B12 and folic acid. Behavioral tests were carried out and blood samples were collected to perform the biochemical profile. Hippocampal tissues were isolated to measure oxidative markers and gene expression. Tissues were also examined histopathologically. Results Memory, anxiety and depressive-like behavior tests, glucometabolic parameters, oxidative stress markers, and gene expression showed significant improvement in all treated groups compared to the untreated diabetic group (P <0.05, for each) except SOD in the IF-treated group (P > 0.05). A significant difference was detected in all studied parameters in the combined treated group compared to the vitamin-treated group and the IF-treated group (P <0.05, for each). Conclusion IF and vitamin supplementation alone or combined can improve the behavioral, biochemical, and histopathological profiles in T2DM with enhanced effects evidenced in combined treatment. In addition, the vitamin-treated group has better effects than the IF-treated group in some metabolic, oxidative stress, and gene expression markers.