Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan; Department for Medical Innovation and Translational Medical Science, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan.
Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan.
Arch Biochem Biophys. 2019 Nov 30;677:108183. doi: 10.1016/j.abb.2019.108183. Epub 2019 Nov 5.
The prostaglandin D metabolite, 15-deoxy-Δ-Prostaglandin J (15d-PGJ), exerts an anti-inflammatory effect through peroxisome proliferator-activated receptor γ (PPARγ)-dependent and -independent anti-inflammatory actions. In the present study, we focused on heme oxygenase-1 (HO-1) induced by 15d-PGJ, and evaluated the effects of enema treatment with 15d-PGJ in the development of intestinal inflammation using a murine colitis model. Acute colitis was induced with dextran sulfate sodium (DSS) in male C57BL/6 mice (8 weeks old) and NF-E2-related factor-2 (Nrf2) deficient mice. Mice were rectally administered 15d-PGJ (1 μM, 0.2 mL: 66.9 ng) daily during DSS administration. Intestinal expression of HO-1 mRNA and protein after rectal administration of 15d-PGJ was evaluated by real-time PCR and western blotting, respectively. A disease activity index (DAI) was determined on a daily basis for each animal, and consisted of a calculated score based on changes in body weight, stool consistency, and intestinal bleeding. Tissue-associated myeloperoxidase (MPO) activity as an index of neutrophil infiltration and mRNA expression levels of TNF-α, IFN-γ, and IL-17A were measured in the colonic mucosa. In addition, we evaluated the effects of co-treatment with a HO-1 inhibitor, zinc protoporphyrin IX (ZnPP), or a specific PPARγ antagonist, GW9662. As a result, rectal administration of 15d-PGJ markedly induced HO-1 protein and mRNA expression in the colonic mucosa. Treatment with 15d-PGJ ameliorated the increase in DAI score and MPO activity and the mRNA expression levels of TNF-α, IFN-γ, and IL-17A after DSS administration. These effects of 15d-PGJ against intestinal inflammation were negated by co-treatment with ZnPP, but not with GW9662. In Nrf2 deficient mice, the rectal administration of 15d-PGJ did not affect colonic HO-1 expression and activity of DSS-induced colitis. These results demonstrate that 15d-PGJ inhibits development of intestinal inflammation in mice via PPAR-independent and Nrf2-HO-1-dependent mechanisms.
前列腺素 D 代谢物 15-脱氧-Δ-前列腺素 J（15d-PGJ）通过过氧化物酶体增殖物激活受体 γ（PPARγ）依赖性和非依赖性抗炎作用发挥抗炎作用。在本研究中，我们专注于 15d-PGJ 诱导的血红素加氧酶-1（HO-1），并使用小鼠结肠炎模型评估了用 15d-PGJ 灌肠治疗对肠道炎症发展的影响。雄性 C57BL/6 小鼠（8 周龄）和 NF-E2 相关因子-2（Nrf2）缺陷小鼠用葡聚糖硫酸钠（DSS）诱导急性结肠炎。在 DSS 给药期间，每天通过直肠给予 15d-PGJ（1μM，0.2mL：66.9ng）。通过实时 PCR 和 Western 印迹分别评估直肠给予 15d-PGJ 后 HO-1 mRNA 和蛋白的肠内表达。每天对每只动物进行疾病活动指数（DAI）的测定，该指数基于体重、粪便稠度和肠道出血的变化计算得出。测量结肠黏膜中髓过氧化物酶（MPO）活性作为中性粒细胞浸润的指标以及 TNF-α、IFN-γ 和 IL-17A 的 mRNA 表达水平。此外，我们评估了 HO-1 抑制剂锌原卟啉 IX（ZnPP）或特定 PPARγ 拮抗剂 GW9662 的共同治疗的效果。结果，直肠给予 15d-PGJ 可显著诱导结肠黏膜中 HO-1 蛋白和 mRNA 的表达。与 DSS 给药后 DAI 评分和 MPO 活性以及 TNF-α、IFN-γ 和 IL-17A 的 mRNA 表达水平的增加相比，15d-PGJ 治疗可改善这些作用。15d-PGJ 对肠道炎症的这些作用被 ZnPP 共同治疗所否定，但 GW9662 不被否定。在 Nrf2 缺陷小鼠中，15d-PGJ 的直肠给药不会影响 DSS 诱导的结肠炎中结肠 HO-1 的表达和活性。这些结果表明，15d-PGJ 通过 PPAR 非依赖性和 Nrf2-HO-1 依赖性机制抑制小鼠肠道炎症的发展。
