Ulcerative colitis (UC) is a kind of the intestinal immune system disorder disease, the exact pathogenesis of UC remains undetermined. Current therapeutic strategies for treating UC mainly relied on nonspecific immune-suppressive therapies. Thus, new therapies are highlight to develop. We have investigated the effects of a kallistatin (Kal)-encoding plasmid in mice acute ulcerative colitis (AUC) by intramuscular electroporation. The model of mice AUC was induced by giving 4% dextran sulfate sodium (DSS) solutions as drinking water 6 days, which was evidenced by Disease activity index (DAI) estimation. Plasmid DNAs were electrotransfered into skeletal muscle, with the best electroporation conditions. Our results suggested that Kal was sufficiently expressed after injection of naked plasmid by intramuscular electroporation. In the Kal gene-transferred group, the colonic myeloperoxidase (MPO) activity, malondialdehyde level and TNF-α content were declined , the superoxide dismutase(SOD)activity was enhanced, and all had significant differences (p＜0.05) compared to those in non-Kal-treated group. The electrotransfed Kal plasmid can express transgene effectively in vivo and this might be a promising approach for treating UC.