Volume 5, Issue 3, Pages 191-198 (December 1998) Dysfunction of antioxidative enzymes in the trinitrobenzenesulfonic acid-induced colitis rat  Toshiya Watanabe, Junichi Fujii, Keiichiro Suzuki, Tsuneko Fujii, Michio Asahi, Ken Matsuoka, Naoyuki Taniguchi  Pathophysiology  Volume 5, Issue 3, Pages 191-198 (December 1998) DOI: 10.1016/S0928-4680(98)00020-0

Fig. 1 SOD and GPx activities in colitis tissues and RASMC after administration of TNBS. (A) SOD activities in colitis tissues (A) and cultured RASMC (B) were measured at 0, 24, and 48 h after TNBS treatment. *, #,+, P<0.001. (C) GPx activities in both colitis tissues and RASMC were measured at 0, 24, and 48 h after TNBS treatment. *, #, P<0.001 compared to controls. Pathophysiology 1998 5, 191-198DOI: (10.1016/S0928-4680(98)00020-0)

Fig. 2 Changes of MnSOD levels in rat colon tissues (A) and RASMC (B) after TNBS administration. Typical data from several experiments were shown. After fractionation of 45 μg proteins by SDS-PAGE, they were blotted onto nitrocellulose membranes. An anti-rat MnSOD antibody was used as the primary antibody. Bands of 25 kDa correspond to MnSOD monomer. MnSOD, 2 μg MnSOD purified from rat liver. Pathophysiology 1998 5, 191-198DOI: (10.1016/S0928-4680(98)00020-0)

Fig. 3 Detection of nitrotyrosine in colitis tissues (A) and RASMC (B). Immunoblot analyses were carried out using anti-nitrotyrosine antibody as the primary antibody. Peroxynitrite-treated MnSOD was used as the positive control. M, prestained molecular weight marker. Pathophysiology 1998 5, 191-198DOI: (10.1016/S0928-4680(98)00020-0)

Fig. 4 Detection of nitrotyrosine in immunoprecipitated MnSOD. MnSOD was immunoprecipitated from 45 μg proteins of TNBS-induced colitis tissues (A, B) and RASMC (C) and then subjected to SDS-PAGE (15%). Antibodies against anti-nitrotyrosine (A, C) and MnSOD (B) were used as the primary antibodies. Pathophysiology 1998 5, 191-198DOI: (10.1016/S0928-4680(98)00020-0)

Fig. 5 Effects of TNBS and/or ascorbate on purified MnSOD, CuZnSOD, and GPx activities. MnSOD (0.1 mg/ml), CuZnSOD (0.1 mg/ml), and GPx (2 mg/ml) were incubated with TNBS (1 mM) and/or ascorbate (1 mM) for 1 h, and then their activities were measured in duplicate at 37°C. Percent of the original activities were shown. Pathophysiology 1998 5, 191-198DOI: (10.1016/S0928-4680(98)00020-0)