title: Sesamin increases heme oxygenase-1 protein in RAW 264.7 macrophages through inhibiting its ubiquitination process.
abstract: Sesamin is a major component in lignans of sesame seed oil, known to possess potent anti-oxidative capacity. In this study, the variation of heme oxygenase (HO)-1, a kind of anti-oxidative enzyme, by sesamin in murine macrophage cell line RAW 264.7 cells was investigated. Lipopolysaccharide (LPS; 10μg/ml) exposure tended to increase HO-1 protein expression. Co-treatment with 100μM sesamin for 12h up-regulated the HO-1 protein level increased by LPS; however, HO-1 mRNA was unaffected. Sesamin delayed the reversal, by the protein synthesis inhibitor cycloheximide (1μM), of the LPS-induced increase of HO-1 protein level. Meanwhile, sesamin suppressed LPS-induced expression of inducible nitric oxide (NO) synthase (iNOS) protein and associated NO release. LPS-induced increase of iNOS protein expression was also reversed by cycloheximide, which was not affected by sesamin, unlike HO-1. To clarify the mechanisms that underlie the up-regulation of HO-1 protein level by sesamin, the human embryonic kidney (HEK) 293T cell line transfected with Flag-tagged HO-1 was used. A proteasome inhibitor, MG-132 (10μM), stabilized HO-1 protein in HEK 293T cells. Co-treatment with sesamin decreased ubiquitinated HO-1 protein accumulation by MG-132. However, sesamin did not affect the proteasome activity. These findings suggest that sesamin disturbs the degradation of HO-1 protein through inhibiting its ubiquitination, resulting in HO-1 protein up-regulation.
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Mizuki
authors.initials: M
authors.lastname: Fukunaga
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan; Department of Pharmacotherapeutics, Graduate School of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan. Electronic address: ohnishi@fupharm.fukuyama-u.ac.jp.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Ohnishi
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Ayano
authors.initials: A
authors.lastname: Shiratsuchi
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Kawakami
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Madoka
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Misato
authors.initials: M
authors.lastname: Motomura
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Kyohei
authors.initials: K
authors.lastname: Egusa
authors.affiliation: Department of Pharmacotherapeutics, Graduate School of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Tomoka
authors.initials: T
authors.lastname: Urasaki
authors.affiliation: Department of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan; Department of Pharmacotherapeutics, Graduate School of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 7290292, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Inoue
mesh_terms: D000818:Animals; D002460:Cell Line; D004149:Dioxoles / Q000494:pharmacology*; D057809:HEK293 Cells; D051547:Heme Oxygenase-1 / Q000096:biosynthesis*; D006801:Humans; D017705:Lignans / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis*; D051379:Mice; D012715:Sesame Oil / Q000494:pharmacology*; D054875:Ubiquitination / Q000187:drug effects* / Q000502:physiology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
chemical_list: D004149:Dioxoles; D017705:Lignans; D008565:Membrane Proteins; D012715:Sesame Oil; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; C054125:sesamin
keywords: Heme oxygenase-1; Macrophage; Sesamin; Ubiquitination