Limits...
Erucin exerts anti-inflammatory properties in murine macrophages and mouse skin: possible mediation through the inhibition of NFκB signaling.

Cho HJ, Lee KW, Park JH - Int J Mol Sci (2013)

Bottom Line: In RAW 264.7 cells, erucin (2.5, 5 μmol/L) inhibited LPS-induced production of nitric oxide and prostaglandin E2.Erucin inhibited LPS-induced degradation of the inhibitor of κBα and translocation of p65 to the nucleus and, subsequently, reduced LPS-induced nuclear factor κB (NFκB) DNA binding activities, as well as the transcriptional activity of NFκB, leading to the decreased expression of NFκB-target genes, including tumor necrosis factor-α, interleukin (IL)-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, as well as transcriptional activity of iNOS and COX-2.These results indicate that erucin exerts a potent anti-inflammatory activity by inhibiting the pro-inflammatory enzymes and cytokines, which may be mediated, at least in part, via the inhibition of NFκB signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Hallym University, Chuncheon 200-702, Korea. jyoon@hallym.ac.kr.

ABSTRACT
Erucin, an isothiocyanate, is a hydrolysis product of glucoerucin found in arugula and has recently been reported to have anti-cancer properties in various cancer cells. In this study, we assessed the anti-inflammatory effects of erucin and the underlying mechanisms, using lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages and 12-O-tetradecanoylphorbol-13-acetate-treated mouse skin. In RAW 264.7 cells, erucin (2.5, 5 μmol/L) inhibited LPS-induced production of nitric oxide and prostaglandin E2. Erucin inhibited LPS-induced degradation of the inhibitor of κBα and translocation of p65 to the nucleus and, subsequently, reduced LPS-induced nuclear factor κB (NFκB) DNA binding activities, as well as the transcriptional activity of NFκB, leading to the decreased expression of NFκB-target genes, including tumor necrosis factor-α, interleukin (IL)-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, as well as transcriptional activity of iNOS and COX-2. In mice, erucin (100, 300 nmoles) treatment significantly inhibited phorbol ester-induced formation of ear edema and expression of iNOS and COX-2 proteins. These results indicate that erucin exerts a potent anti-inflammatory activity by inhibiting the pro-inflammatory enzymes and cytokines, which may be mediated, at least in part, via the inhibition of NFκB signaling.

Show MeSH

Related in: MedlinePlus

Erucin inhibits LPS-induced activation of NFκB signaling in RAW 264.7 cells. (A,B) RAW 264.7 cells were treated with various concentrations of erucin for 30 min. LPS was then added and incubated for another 20 min. (A) Total cell lysates were subjected to Western blotting with their relevant antibodies; (B) Cytosolic fractions and nuclear extracts were prepared for Western blotting with an anti-p65 antibody and Electrophoretic Mobility Shift Assay (EMSA). The relative abundance of each band was quantified, and the control levels were set at 100%. The adjusted mean ± SEM (n = 3) of each band is shown above each blot; (C) RAW 264.7 cells were transfected with NFκB-luciferase reporter plasmid. The transfected cells were treated with various concentrations of erucin in the presence of LPS. Cell lysates were prepared to determine luciferase activity. Each bar represents the mean ± SEM (n = 4). Means without a common letter differ (p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3821631&req=5

f4-ijms-14-20564: Erucin inhibits LPS-induced activation of NFκB signaling in RAW 264.7 cells. (A,B) RAW 264.7 cells were treated with various concentrations of erucin for 30 min. LPS was then added and incubated for another 20 min. (A) Total cell lysates were subjected to Western blotting with their relevant antibodies; (B) Cytosolic fractions and nuclear extracts were prepared for Western blotting with an anti-p65 antibody and Electrophoretic Mobility Shift Assay (EMSA). The relative abundance of each band was quantified, and the control levels were set at 100%. The adjusted mean ± SEM (n = 3) of each band is shown above each blot; (C) RAW 264.7 cells were transfected with NFκB-luciferase reporter plasmid. The transfected cells were treated with various concentrations of erucin in the presence of LPS. Cell lysates were prepared to determine luciferase activity. Each bar represents the mean ± SEM (n = 4). Means without a common letter differ (p < 0.05).

Mentions: As NFκB regulates the expression of genes encoding iNOS, COX-2, TNF-α, IL-6 and IL-1β [18,19], we next determined whether erucin inhibits NFκB signaling. Upon stimulation, inhibitors of NFκB (IκB)-α are phosphorylated and degraded, and the NFκB proteins in the cytoplasm are liberated from IκB and translocated to the nucleus, where they bind to the promoter regions of NFκB-responsive genes [15]. LPS treatment reduced the levels of IκB-α, and erucin suppressed the LPS-induced reduction in IκB-α (Figure 4A). The levels of cytosolic p65 protein were reduced by LPS treatment, which was suppressed by erucin, whereas the levels of nuclear p65 were markedly increased by LPS, and this increase was suppressed by erucin treatment. Additionally, NFκB DNA binding was markedly increased by LPS, which was suppressed by erucin pre-treatment (Figure 4B). Furthermore, erucin inhibited the transcriptional activity of NFκB in LPS-stimulated cells (Figure 4C).


Erucin exerts anti-inflammatory properties in murine macrophages and mouse skin: possible mediation through the inhibition of NFκB signaling.

Cho HJ, Lee KW, Park JH - Int J Mol Sci (2013)

Erucin inhibits LPS-induced activation of NFκB signaling in RAW 264.7 cells. (A,B) RAW 264.7 cells were treated with various concentrations of erucin for 30 min. LPS was then added and incubated for another 20 min. (A) Total cell lysates were subjected to Western blotting with their relevant antibodies; (B) Cytosolic fractions and nuclear extracts were prepared for Western blotting with an anti-p65 antibody and Electrophoretic Mobility Shift Assay (EMSA). The relative abundance of each band was quantified, and the control levels were set at 100%. The adjusted mean ± SEM (n = 3) of each band is shown above each blot; (C) RAW 264.7 cells were transfected with NFκB-luciferase reporter plasmid. The transfected cells were treated with various concentrations of erucin in the presence of LPS. Cell lysates were prepared to determine luciferase activity. Each bar represents the mean ± SEM (n = 4). Means without a common letter differ (p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3821631&req=5

f4-ijms-14-20564: Erucin inhibits LPS-induced activation of NFκB signaling in RAW 264.7 cells. (A,B) RAW 264.7 cells were treated with various concentrations of erucin for 30 min. LPS was then added and incubated for another 20 min. (A) Total cell lysates were subjected to Western blotting with their relevant antibodies; (B) Cytosolic fractions and nuclear extracts were prepared for Western blotting with an anti-p65 antibody and Electrophoretic Mobility Shift Assay (EMSA). The relative abundance of each band was quantified, and the control levels were set at 100%. The adjusted mean ± SEM (n = 3) of each band is shown above each blot; (C) RAW 264.7 cells were transfected with NFκB-luciferase reporter plasmid. The transfected cells were treated with various concentrations of erucin in the presence of LPS. Cell lysates were prepared to determine luciferase activity. Each bar represents the mean ± SEM (n = 4). Means without a common letter differ (p < 0.05).
Mentions: As NFκB regulates the expression of genes encoding iNOS, COX-2, TNF-α, IL-6 and IL-1β [18,19], we next determined whether erucin inhibits NFκB signaling. Upon stimulation, inhibitors of NFκB (IκB)-α are phosphorylated and degraded, and the NFκB proteins in the cytoplasm are liberated from IκB and translocated to the nucleus, where they bind to the promoter regions of NFκB-responsive genes [15]. LPS treatment reduced the levels of IκB-α, and erucin suppressed the LPS-induced reduction in IκB-α (Figure 4A). The levels of cytosolic p65 protein were reduced by LPS treatment, which was suppressed by erucin, whereas the levels of nuclear p65 were markedly increased by LPS, and this increase was suppressed by erucin treatment. Additionally, NFκB DNA binding was markedly increased by LPS, which was suppressed by erucin pre-treatment (Figure 4B). Furthermore, erucin inhibited the transcriptional activity of NFκB in LPS-stimulated cells (Figure 4C).

Bottom Line: In RAW 264.7 cells, erucin (2.5, 5 μmol/L) inhibited LPS-induced production of nitric oxide and prostaglandin E2.Erucin inhibited LPS-induced degradation of the inhibitor of κBα and translocation of p65 to the nucleus and, subsequently, reduced LPS-induced nuclear factor κB (NFκB) DNA binding activities, as well as the transcriptional activity of NFκB, leading to the decreased expression of NFκB-target genes, including tumor necrosis factor-α, interleukin (IL)-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, as well as transcriptional activity of iNOS and COX-2.These results indicate that erucin exerts a potent anti-inflammatory activity by inhibiting the pro-inflammatory enzymes and cytokines, which may be mediated, at least in part, via the inhibition of NFκB signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Hallym University, Chuncheon 200-702, Korea. jyoon@hallym.ac.kr.

ABSTRACT
Erucin, an isothiocyanate, is a hydrolysis product of glucoerucin found in arugula and has recently been reported to have anti-cancer properties in various cancer cells. In this study, we assessed the anti-inflammatory effects of erucin and the underlying mechanisms, using lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages and 12-O-tetradecanoylphorbol-13-acetate-treated mouse skin. In RAW 264.7 cells, erucin (2.5, 5 μmol/L) inhibited LPS-induced production of nitric oxide and prostaglandin E2. Erucin inhibited LPS-induced degradation of the inhibitor of κBα and translocation of p65 to the nucleus and, subsequently, reduced LPS-induced nuclear factor κB (NFκB) DNA binding activities, as well as the transcriptional activity of NFκB, leading to the decreased expression of NFκB-target genes, including tumor necrosis factor-α, interleukin (IL)-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, as well as transcriptional activity of iNOS and COX-2. In mice, erucin (100, 300 nmoles) treatment significantly inhibited phorbol ester-induced formation of ear edema and expression of iNOS and COX-2 proteins. These results indicate that erucin exerts a potent anti-inflammatory activity by inhibiting the pro-inflammatory enzymes and cytokines, which may be mediated, at least in part, via the inhibition of NFκB signaling.

Show MeSH
Related in: MedlinePlus