Resolvin E1, an endogenous lipid mediator derived from eicosapentaenoic acid, prevents dextran sulfate sodium-induced colitis - PubMed (original) (raw)

Masaru Yoshida, Makoto Arita, Yosuke Nishitani, Shin Nishiumi, Atsuhiro Masuda, Shigeto Mizuno, Tetsuya Takagawa, Yoshinori Morita, Hiromu Kutsumi, Hideto Inokuchi, Charles N Serhan, Richard S Blumberg, Takeshi Azuma

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Resolvin E1, an endogenous lipid mediator derived from eicosapentaenoic acid, prevents dextran sulfate sodium-induced colitis

Tsukasa Ishida et al. Inflamm Bowel Dis. 2010 Jan.

Abstract

Background: Resolvin E1 (RvE1), an endogenous lipid mediator derived from eicosapentaenoic acid, has been identified in local inflammation during the healing stage. RvE1 reduces inflammation in several types of animal models including peritonitis and retinopathy and blocks human neutrophil transendothelial cell migration. The RvE1 receptor ChemR23 is expressed on myeloid cells such as macrophages and dendritic cells. The aim of this study was to determine whether RvE1 regulates colonic inflammation when the innate immune response of macrophages plays a key role in pathogenesis and tissue damage.

Methods: The RvE1 receptor ChemR23 was expressed in mouse peritoneal macrophages as defined by flow cytometry. Peritoneal macrophages were pretreated with RvE1, followed by lipopolysaccharide stimulation, whereupon transcriptional levels of proinflammatory cytokines were analyzed.

Results: RvE1 treatment led to inhibition of proinflammatory cytokines including TNF-alpha and IL-12p40. In HEK293 cells, pretreatment with RvE1 inhibited TNF-alpha-induced nuclear translocation of NF-kappaB in a ChemR23-dependent manner. These results suggested that RvE1 could regulate proinflammatory responses of macrophages expressing ChemR23. Therefore, we investigated the beneficial effects of RvE1 in dextran sulfate sodium-induced colitis. RvE1 treatment led to amelioration of colonic inflammation.

Conclusions: These results indicate that RvE1 suppresses proinflammatory responses of macrophages. RvE1 and its receptor may therefore be useful as therapeutic targets in the treatment of human inflammatory bowel disease and other inflammatory disorders.

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Figures

Figure 1

Figure 1. ChemR23 expresses on macrophages, and regulates proinflammatory cytokines

(A) Anti-mChemR23 mAb and FITC labeled anti-mCD11b mAb staining of primary mouse peritoneal macrophages. (B) Primary peritoneal macrophages were pretreated with RvE1 (100 ng/ml) for 4 hours followed by stimulation with LPS (100 ng/ml), and cytokine mRNA was analyzed by quantitative RT-PCR. Data are shown as the mean ± S.E. (n=3).

Figure 2

Figure 2. RvE1 inhibits the nuclear translocation of NF-κB via ChemR23

(A) Staining of Human ChemR23 stably-expressed HEK293 cells by anti-human ChemR23 mAb. (B) HEK293 wild type cells or ChemR23 transfected cells were incubated with 100 ng/ml RvE1 for 2 hours followed by treatment with 4 ng/ml TNF-α for 1 hour, and then subjected to immunocytochemistry. (C) HEK293 wild type cells or ChemR23 transfected cells were incubated with 100 ng/ml RvE1 for 2 hours followed by treatment with 4 ng/ml TNF-α for 1 hour. The nuclear proteins were prepared and analyzed for NF-κB p65 by Western blotting analysis. The band density of NF-κB was quantified, and the values relative to a vehicle control are shown. Data are shown as the mean ± S.E. (n=3).

Figure 3

Figure 3. RvE1 dramatically attenuates DSS-induced colitis

(A) Wasting disease as measured by body weight loss in mice with DSS-induced colitis is improved by RvE1 treatment. Open circles, DSS alone; filled circles, DSS treated with RvE1 (1 µg per mouse) daily for five days (day0–5). *, P< 0.05, **, P< 0.01 (_n_=6). (B) Length of the inflamed colons of mice treated with either DSS alone or DSS plus RvE1. *P< 0.01, compared with DSS alone. (C) Colon sections of mice exposed to drinking water only (Left), day 8 after DSS-induced colitis (Center), and RvE1-treated mice (Right).(D) Histological scores of the colon of mice receiving DSS alone or DSS plus RvE1. *P<0.05, compared with DSS alone. Data are shown as the mean ± S.E. (n=6).

Figure 4

Figure 4. RvE1 reduces NF-κB phosphorylation and NF-κB-dependent proinflammatory mediators in DSS-induced colitis

(A) Immunohistchemical analysis of the effects of RvE1 on NF-κB activation (NF-κB p65 Phosphorylation at Ser276) in distal colon of mice receiving water only, DSS only, and DSS plus RvE1. Brown staining indicates cells positive for Ser276 phosphorylation of NF-κB p65. (magnification ×200: upper side, and ×400: lower side) (B) Staining was scored by counting the number of Ser276-phosphorylated NF-κB p65-positive cells (colonic epithelial cells; CEC: open bars, and lamina propria lymphocytes; LPL: shaded bars) per visualized high-power field (magnification × 400) in distal colon of mice receiving water only, DSS only, and DSS plus RvE1. (C) Quantitative real-time PCR analysis of mRNA expression of inflammatory mediators in colons obtained on day 8 from control mice (white column), mice treated with DSS alone (black column) or mice receiving DSS plus RvE1 (gray column). *, P<0.05, compared with DSS alone. Data are shown as the mean ± S.E. (n=6).

Figure 5

Figure 5. ChemR23 mRNA increases during LPS-stimulation and DSS induced colitis

(A) ChemR23 expression on peritoneal macrophages after 100 ng/ml LPS stimulation for 24 hours (black column) in comparison to no stimulation (white column). (B) ChemR23 expression in colon tissues of DSS induced colitis (black column) in comparison to control mice (white column). Data are shown as the mean ± S.E. (n=6).

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