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Fisetin Suppresses Macrophage-Mediated Inflammatory Responses by Blockade of Src and Syk.

Kim JH, Kim MY, Kim JH, Cho JY - Biomol Ther (Seoul) (2015)

Bottom Line: In agreement, the upstream phosphorylation events for NF-κB activation, composed of Src, Syk, and IκBα, were also reduced by fisetin.The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin.Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746.

ABSTRACT
Flavonoids, such as fisetin (3,7,3',4'-tetrahydroxyflavone), are plant secondary metabolites. It has been reported that fisetin is able to perform numerous pharmacological roles including anti-inflammatory, anti-microbial, and anti-cancer activities; however, the exact anti-inflammatory mechanism of fisetin is not understood. In this study, the pharmacological action modes of fisetin in lipopolysaccharide (LPS)-stimulated macrophage-like cells were elucidated by using immunoblotting analysis, kinase assays, and an overexpression strategy. Fisetin diminished the release of nitric oxide (NO) and reduced the mRNA levels of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, and cyclooxygenase (COX)-2 in LPS-stimulated RAW264.7 cells without displaying cytotoxicity. This compound also blocked the nuclear translocation of p65/nuclear factor (NF)-κB. In agreement, the upstream phosphorylation events for NF-κB activation, composed of Src, Syk, and IκBα, were also reduced by fisetin. The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin. Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

No MeSH data available.


Related in: MedlinePlus

The effect of fisetin on transcriptional activation in LPS-stimulated RAW264.7 cells. (A and B) The mRNA levels of iNOS, COX-2, and TNF-α expressed in LPS (1 μg/mL)-treated RAW264.7 cells in the presence or absence of fisetin (20 and 30 μM) were measured by RT-PCR (A) or real-time PCR (B). (C) The nuclear levels of p65 and p50 in RAW264.7 cells treated with LPS (1 μg/mL) in the presence or absence of fisetin (30 VM) were analyzed by immunoblotting analysis. Data (B) are expressed as the mean ± SD of experiments, which were performed with six samples. **p<0.01 compared to normal or control groups.
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f3-bt-23-414: The effect of fisetin on transcriptional activation in LPS-stimulated RAW264.7 cells. (A and B) The mRNA levels of iNOS, COX-2, and TNF-α expressed in LPS (1 μg/mL)-treated RAW264.7 cells in the presence or absence of fisetin (20 and 30 μM) were measured by RT-PCR (A) or real-time PCR (B). (C) The nuclear levels of p65 and p50 in RAW264.7 cells treated with LPS (1 μg/mL) in the presence or absence of fisetin (30 VM) were analyzed by immunoblotting analysis. Data (B) are expressed as the mean ± SD of experiments, which were performed with six samples. **p<0.01 compared to normal or control groups.

Mentions: To check whether the anti-inflammatory effect of fisetin occurs at the transcriptional level, we determined the mRNA expression levels of inflammatory genes such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-α (TNF-α). As expected, the expression of inflammatory mediator genes was suppressed in cells treated with either 20 or 30 μM of fisetin according to both semi-quantitative (Fig. 3A) and real-time RT-PCR (Fig. 3B) analyses. We also examined transcription factor levels in nuclear extracts by immunoblotting analysis and found that fisetin treatment was capable of inhibiting the nuclear translocation of p65/NF-κB at 60 min but not of p50/NF-κB (Fig. 3C).


Fisetin Suppresses Macrophage-Mediated Inflammatory Responses by Blockade of Src and Syk.

Kim JH, Kim MY, Kim JH, Cho JY - Biomol Ther (Seoul) (2015)

The effect of fisetin on transcriptional activation in LPS-stimulated RAW264.7 cells. (A and B) The mRNA levels of iNOS, COX-2, and TNF-α expressed in LPS (1 μg/mL)-treated RAW264.7 cells in the presence or absence of fisetin (20 and 30 μM) were measured by RT-PCR (A) or real-time PCR (B). (C) The nuclear levels of p65 and p50 in RAW264.7 cells treated with LPS (1 μg/mL) in the presence or absence of fisetin (30 VM) were analyzed by immunoblotting analysis. Data (B) are expressed as the mean ± SD of experiments, which were performed with six samples. **p<0.01 compared to normal or control groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556200&req=5

f3-bt-23-414: The effect of fisetin on transcriptional activation in LPS-stimulated RAW264.7 cells. (A and B) The mRNA levels of iNOS, COX-2, and TNF-α expressed in LPS (1 μg/mL)-treated RAW264.7 cells in the presence or absence of fisetin (20 and 30 μM) were measured by RT-PCR (A) or real-time PCR (B). (C) The nuclear levels of p65 and p50 in RAW264.7 cells treated with LPS (1 μg/mL) in the presence or absence of fisetin (30 VM) were analyzed by immunoblotting analysis. Data (B) are expressed as the mean ± SD of experiments, which were performed with six samples. **p<0.01 compared to normal or control groups.
Mentions: To check whether the anti-inflammatory effect of fisetin occurs at the transcriptional level, we determined the mRNA expression levels of inflammatory genes such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-α (TNF-α). As expected, the expression of inflammatory mediator genes was suppressed in cells treated with either 20 or 30 μM of fisetin according to both semi-quantitative (Fig. 3A) and real-time RT-PCR (Fig. 3B) analyses. We also examined transcription factor levels in nuclear extracts by immunoblotting analysis and found that fisetin treatment was capable of inhibiting the nuclear translocation of p65/NF-κB at 60 min but not of p50/NF-κB (Fig. 3C).

Bottom Line: In agreement, the upstream phosphorylation events for NF-κB activation, composed of Src, Syk, and IκBα, were also reduced by fisetin.The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin.Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746.

ABSTRACT
Flavonoids, such as fisetin (3,7,3',4'-tetrahydroxyflavone), are plant secondary metabolites. It has been reported that fisetin is able to perform numerous pharmacological roles including anti-inflammatory, anti-microbial, and anti-cancer activities; however, the exact anti-inflammatory mechanism of fisetin is not understood. In this study, the pharmacological action modes of fisetin in lipopolysaccharide (LPS)-stimulated macrophage-like cells were elucidated by using immunoblotting analysis, kinase assays, and an overexpression strategy. Fisetin diminished the release of nitric oxide (NO) and reduced the mRNA levels of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, and cyclooxygenase (COX)-2 in LPS-stimulated RAW264.7 cells without displaying cytotoxicity. This compound also blocked the nuclear translocation of p65/nuclear factor (NF)-κB. In agreement, the upstream phosphorylation events for NF-κB activation, composed of Src, Syk, and IκBα, were also reduced by fisetin. The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin. Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

No MeSH data available.


Related in: MedlinePlus