The present study was designed to investigate the protective effect of hesperidin extract on some blood and biochemical profile in addition interleukin-1 (IL-1) and zonula occluding -1(ZO-1) gene expression of colon tissue in acetic acid-induced ulcerative colitis in rats. Forty rats were divided into four equal groups. Group I (c) group (control normal group) was received no drugs. Group II (AA) group (Acetic acid induced colitis group) rats were administrated with 2 ml of acetic acid (AA) 4% intracolonic as a single dose. Group III (HMC) group (Hesperidin methyl chalone group) rats were administrated hesperidin orally (200 mg/kg, b.wt/day) for 14 days. Group IV (HMC+AA) group (Hesperidin protected + acetic acid 4% induce colitis group) rats were administrated orally (200 mg/kg, b.wt/day) for 14 days and at the 11th day were administrated intracolonic with 2 ml AA 4% as a single dose. The obtained results showed that significant decrease in neutrophils to lymphocytes ratio (N/LR) and increase in lymphocytes to monocytes ratio (L/MR) in (HMC+AA) group when compared with (AA) group. In addition, ALP in serum was significantly decrease with significant down-regulation in IL-1ß, inducible nitric oxide (iNOS), cyclooxygenase-2 (COX2) and ZO-1 gene expression level in colon tissue of (HMC+AA) group when compared with (AA) group. The results revealed that hesperidin has protective, anti-inflammatory and antioxidant effects in experimental model of ulcerative colitis induced in rats.