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Moringa oleifera Flower Extract Suppresses the Activation of Inflammatory Mediators in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages via NF-κB Pathway.

Tan WS, Arulselvan P, Karthivashan G, Fakurazi S - Mediators Inflamm. (2015)

Bottom Line: However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB) in a concentration dependent manner (100 μg/mL and 200 μg/mL).Conclusion.These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator's production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.

ABSTRACT
Aim of Study. Moringa oleifera Lam. (M. oleifera) possess highest concentration of antioxidant bioactive compounds and is anticipated to be used as an alternative medicine for inflammation. In the present study, we investigated the anti-inflammatory activity of 80% hydroethanolic extract of M. oleifera flower on proinflammatory mediators and cytokines produced in lipopolysaccharide- (LPS-) induced RAW 264.7 macrophages. Materials and Methods. Cell cytotoxicity was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide (NO) production was quantified through Griess reaction while proinflammatory cytokines and other key inflammatory markers were assessed through enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results. Hydroethanolic extract of M. oleifera flower significantly suppressed the secretion and expression of NO, prostaglandin E2 (PGE2), interleukin- (IL-) 6, IL-1β, tumor necrosis factor-alpha (TNF-α), nuclear factor-kappa B (NF-κB), inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2). However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB) in a concentration dependent manner (100 μg/mL and 200 μg/mL). Conclusion. These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator's production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway.

No MeSH data available.


Related in: MedlinePlus

(a) Anti-inflammatory effect of 80% hydroethanolic M. oleifera bioactive flower extract on the expression of NF-κB, IκB-α, iNOS, and COX-2 in LPS-induced RAW 264.7 macrophages. A density of 1 × 106 cells/well of macrophages in the presence or absence of LPS were seeded in 6-well plate and treated with indicated concentrations of flower extract for 24 h. The protein of cells was collected through RIPA buffer and analysed by Western blotting. β-actin acts as a loading control and also standard for target proteins in quantitative determination. (b) Densitometry analysis results of the effect of M. oleifera flower extract on proteins expression. ###p < 0.001 and ##p < 0.01 were LPS-treated group versus control; ∗∗∗p < 0.001: treated group significantly different from LPS-treated group. Control: basal level of cytokines released without LPS induction. The data are presented as mean ± SD of three independent experiments.
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fig6: (a) Anti-inflammatory effect of 80% hydroethanolic M. oleifera bioactive flower extract on the expression of NF-κB, IκB-α, iNOS, and COX-2 in LPS-induced RAW 264.7 macrophages. A density of 1 × 106 cells/well of macrophages in the presence or absence of LPS were seeded in 6-well plate and treated with indicated concentrations of flower extract for 24 h. The protein of cells was collected through RIPA buffer and analysed by Western blotting. β-actin acts as a loading control and also standard for target proteins in quantitative determination. (b) Densitometry analysis results of the effect of M. oleifera flower extract on proteins expression. ###p < 0.001 and ##p < 0.01 were LPS-treated group versus control; ∗∗∗p < 0.001: treated group significantly different from LPS-treated group. Control: basal level of cytokines released without LPS induction. The data are presented as mean ± SD of three independent experiments.

Mentions: Immunoblotting was conducted to evaluate the expression of inflammatory mediators which included NF-κB, IκB-α, iNOS, and COX-2 in LPS-stimulated RAW 264.7 macrophages treated with the 80% hydroethanolic M. oleifera flower extract at concentrations 100 and 200 μg/mL. As illustrated in Figure 6, the NF-κB, iNOS, and COX-2 target markers are significantly expressed in the LPS-treated group compared to the control untreated group. However, the treatment of M. oleifera flower extract concentration dependently downregulated the target molecule expressions in LPS-stimulated macrophages. On the other hand, IκB-α expression is increased with the presence of flower extract.


Moringa oleifera Flower Extract Suppresses the Activation of Inflammatory Mediators in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages via NF-κB Pathway.

Tan WS, Arulselvan P, Karthivashan G, Fakurazi S - Mediators Inflamm. (2015)

(a) Anti-inflammatory effect of 80% hydroethanolic M. oleifera bioactive flower extract on the expression of NF-κB, IκB-α, iNOS, and COX-2 in LPS-induced RAW 264.7 macrophages. A density of 1 × 106 cells/well of macrophages in the presence or absence of LPS were seeded in 6-well plate and treated with indicated concentrations of flower extract for 24 h. The protein of cells was collected through RIPA buffer and analysed by Western blotting. β-actin acts as a loading control and also standard for target proteins in quantitative determination. (b) Densitometry analysis results of the effect of M. oleifera flower extract on proteins expression. ###p < 0.001 and ##p < 0.01 were LPS-treated group versus control; ∗∗∗p < 0.001: treated group significantly different from LPS-treated group. Control: basal level of cytokines released without LPS induction. The data are presented as mean ± SD of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig6: (a) Anti-inflammatory effect of 80% hydroethanolic M. oleifera bioactive flower extract on the expression of NF-κB, IκB-α, iNOS, and COX-2 in LPS-induced RAW 264.7 macrophages. A density of 1 × 106 cells/well of macrophages in the presence or absence of LPS were seeded in 6-well plate and treated with indicated concentrations of flower extract for 24 h. The protein of cells was collected through RIPA buffer and analysed by Western blotting. β-actin acts as a loading control and also standard for target proteins in quantitative determination. (b) Densitometry analysis results of the effect of M. oleifera flower extract on proteins expression. ###p < 0.001 and ##p < 0.01 were LPS-treated group versus control; ∗∗∗p < 0.001: treated group significantly different from LPS-treated group. Control: basal level of cytokines released without LPS induction. The data are presented as mean ± SD of three independent experiments.
Mentions: Immunoblotting was conducted to evaluate the expression of inflammatory mediators which included NF-κB, IκB-α, iNOS, and COX-2 in LPS-stimulated RAW 264.7 macrophages treated with the 80% hydroethanolic M. oleifera flower extract at concentrations 100 and 200 μg/mL. As illustrated in Figure 6, the NF-κB, iNOS, and COX-2 target markers are significantly expressed in the LPS-treated group compared to the control untreated group. However, the treatment of M. oleifera flower extract concentration dependently downregulated the target molecule expressions in LPS-stimulated macrophages. On the other hand, IκB-α expression is increased with the presence of flower extract.

Bottom Line: However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB) in a concentration dependent manner (100 μg/mL and 200 μg/mL).Conclusion.These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator's production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.

ABSTRACT
Aim of Study. Moringa oleifera Lam. (M. oleifera) possess highest concentration of antioxidant bioactive compounds and is anticipated to be used as an alternative medicine for inflammation. In the present study, we investigated the anti-inflammatory activity of 80% hydroethanolic extract of M. oleifera flower on proinflammatory mediators and cytokines produced in lipopolysaccharide- (LPS-) induced RAW 264.7 macrophages. Materials and Methods. Cell cytotoxicity was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide (NO) production was quantified through Griess reaction while proinflammatory cytokines and other key inflammatory markers were assessed through enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results. Hydroethanolic extract of M. oleifera flower significantly suppressed the secretion and expression of NO, prostaglandin E2 (PGE2), interleukin- (IL-) 6, IL-1β, tumor necrosis factor-alpha (TNF-α), nuclear factor-kappa B (NF-κB), inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2). However, it significantly increased the production of IL-10 and IκB-α (inhibitor of κB) in a concentration dependent manner (100 μg/mL and 200 μg/mL). Conclusion. These results suggest that 80% hydroethanolic extract of M. oleifera flower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator's production in LPS-stimulated macrophages through preventing degradation of IκB-α in NF-κB signaling pathway.

No MeSH data available.


Related in: MedlinePlus