Diabetes comorbidity in ulcerative colitis (UC) has relevant clinical and therapeutic implications. The link between hyperglycemia and intestinal barrier function with respect to infection and inflammation consequences exists in diabetes. The present study was designed to decipher the molecular mechanisms associated with Type 1 Diabetes mellitus and the UC in both male and female BALB/c mice. Dextran sulfate sodium (DSS; 2.5%w/v) dissolved in drinking water was given for three cycles (each cycle; 7 days) with 7 days recovery period in-between to both male and female BALB/c mice. At the first recovery period, Streptozotocin (40 mg/kg; i.p.) was administered for 5 consecutive days in the case of male BALB/c mice; whereas the same procedure was repeated at the beginning of each recovery period in female animals. In the DSS + DB group of male animals, disease activity index, myeloperoxidase activity, nitrite level, plasma lipopolysaccharides, interleukin-1β, histological score, % fibrotic area, % TUNEL positive cells were significantly increased. Furthermore, protein expression of phosphorylated nuclear factor kappa light chain enhancer of activated B cells (pNFκB65), proliferating cell nuclear antigen, interleukin-6, apoptosis-associated speck-like protein containing a caspase-recruitment domain, and cysteine-containing aspartate-specific proteases-1 (caspase-1) significantly increased in the DSS + DB group of male animals as compared to female. The present study findings proved that hyperglycemic conditions exacerbated the pathological conditions in UC of male animals; whereas milder conditions developed in females.