This work investigated the physiochemical characteristics and preventive effects of purified pectin (H121, L13 and L102) with different esterification degrees on dextran sulfate sodium (DSS)-induced colitis in mice. Three doses of each type of pectin were administered to C57BL/6J mice for 7 days before the DSS treatment, with dextran and mesalazine as positive controls. Results showed that pathological factors including the body weight, the disease activity index (DAI), the colonic weight/length ratio and the organ index of the spleen were improved with pre-intervention of a high dose of L13 or L102. Further studies showed that administration of a low dose of L13, low dose and medium dose of L102 or dextran improved intestinal permeability and tight junction function in colitis mice. Treatments of L13 of all doses and L102 of a high dose downregulated the oxidative stress-associated factors, while L102 of a low dose and H121 ameliorated the inflammatory cytokine production in serum and the colon. The above results suggested that pectin could attenuate DSS-induced intestinal epithelial injury, inflammation and oxidative stress. Specifically, compared to high esterified pectin, low esterified pectin displayed better protective effects in acute colitis mice.