Fig 6. MES₅₅₀₀ induces production of H₂O₂, which is partly responsible for its anti-inflammatory effect.

(A) H₂O₂ concentration in MES_CFM. (B) IL-2 mRNA level in Jurkat T cells treated with H₂O₂ and stimulated with PMA/Io. (C) Immunoblots of nuclear and cytosolic fractions of Jurkat cells incubated with catalase-treated H₂O₂ and stimulated with PMA/Io. (D-F) Jurkat T cells were treated with MES_CFM (D) or MES₅₅₀₀ (E, F) with or without catalase (CAT), and stimulated with PMA/Io. Total RNA was analyzed for the indicated genes. (G, H) Immunoblots of nuclear and cytosolic fractions of cells treated with MES_CFM (G) or MES₅₅₀₀ (H) with or without catalase, and stimulated with PMA/Io. (I) Jurkat T cells were treated with MES₅₅₀₀ or H₂O₂ for 10 min and medium was changed. After 24 hr, LDH leakage from cells was measured. (B, D-F) Data were normalized to the level of GAPDH mRNA (internal control). Data are presented as mean ± S.D. (n = 3 per group). (A, F, I) *P<0.05, ***P<0.001 assessed by Wilcoxon test. (B, D, E) *P<0.05, **P<0.01, ***P<0.001 assessed by Tukey-Kramer test. (n.s.; not significant). (J) BALB/c mice (13 weeks old) were treated with MES₅₅₀₀ for 30 min, and serum was obtained to measure H₂O₂ concentration. Data are presented as mean ± S.D. (n = 4 for control group and n = 5 for MES₅₅₀₀-treated group) assessed by Wilcoxon test. The data shown are representative of 2 or more independent experiments.