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Effect of Sulforaphane on NOD2 via NF-κB: implications for Crohn's disease.

Folkard DL, Marlow G, Mithen RF, Ferguson LR - J Inflamm (Lond) (2015)

Bottom Line: These results raise the possibility that PRR-mediated inflammation could be suppressed by specific dietary bioactives.The cells were co-treated with sulforaphane and MDP and secreted alkaline phosphatase (SEAP) production was determined.These results demonstrate that the anti-inflammatory role of sulforaphane is not restricted to LPS-induced inflammatory signalling.

View Article: PubMed Central - PubMed

Affiliation: Food and Health Programme, Institute of Food Research, Norwich Research Park, Norwich, UK.

ABSTRACT

Background: Sulforaphane has well established anti-cancer properties and more recently anti-inflammatory properties have also been determined. Sulforaphane has been shown to inhibit PRR-mediated pro-inflammatory signalling by either directly targeting the receptor or their downstream signalling molecules such as the transcription factor, NF-κB. These results raise the possibility that PRR-mediated inflammation could be suppressed by specific dietary bioactives. We examined whether sulforaphane could suppress NF-κB via the NOD2 pathway.

Methods: Human embryonic kidney 293T (HEK293T) cells were stably transfected with NOD2 variants and the NF-κB reporter, pNifty2-SEAP. The cells were co-treated with sulforaphane and MDP and secreted alkaline phosphatase (SEAP) production was determined.

Results: We found that sulforaphane was able to significantly suppress the ligand-induced NF-κB activity at physiologically relevant concentrations, achievable via the consumption of broccoli within the diet.

Conclusions: These results demonstrate that the anti-inflammatory role of sulforaphane is not restricted to LPS-induced inflammatory signalling. These data add to the growing evidence that PRR activation can be inhibited by specific phytochemicals and thus suggests that diet could be a way of controlling inflammation. This is particularly important for a disease like Crohn's disease where diet can play a key role in relieving or exacerbating symptoms.

No MeSH data available.


Related in: MedlinePlus

L- and D, L-SFN are able to significantly suppress MDP-induced SEAP activity in NOD2 G908R cells. NOD2 G908R cells were treated for 24 h with 0.2 μg/ml MDP in the presence or absence of L- (A) or D, L-SFN (B). Media was collected and added to QUANTI-Blue™ solution and left at room temperature for 5-15 min before being measured using a spectrophotometer at 650 nm. Data shown = mean ± SD, data representative of three independent experiments. + p < 0.05, ++ p < 0.01, +++ p < 0.001 comparison to 0 μM –MDP. *p < 0.05, **p < 0.01, ***p < 0.001, comparison to 0 μM + MDP.
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Fig5: L- and D, L-SFN are able to significantly suppress MDP-induced SEAP activity in NOD2 G908R cells. NOD2 G908R cells were treated for 24 h with 0.2 μg/ml MDP in the presence or absence of L- (A) or D, L-SFN (B). Media was collected and added to QUANTI-Blue™ solution and left at room temperature for 5-15 min before being measured using a spectrophotometer at 650 nm. Data shown = mean ± SD, data representative of three independent experiments. + p < 0.05, ++ p < 0.01, +++ p < 0.001 comparison to 0 μM –MDP. *p < 0.05, **p < 0.01, ***p < 0.001, comparison to 0 μM + MDP.

Mentions: We subsequently confirmed if there was a differential effect based on SFN isomer in the NOD2 WT cells (Figure 4). The only significant difference was seen at 10 μM SFN stimulated with 0.2 μg/ml MDP (Figure 4). This was not as a result of reduced viability with D, L-SFN (data not shown). Finally we examined if the same suppression was true in NOD2 G908R cells. Similarly to the NOD2 WT cells, both SFN isomers showed significant reductions at nearly all the concentrations used (Figure 5). No significant differences were found between the suppression of MDP-induced NF-κB activity when using the L- or the D, L-SFN forms (Figure 6).Figure 4


Effect of Sulforaphane on NOD2 via NF-κB: implications for Crohn's disease.

Folkard DL, Marlow G, Mithen RF, Ferguson LR - J Inflamm (Lond) (2015)

L- and D, L-SFN are able to significantly suppress MDP-induced SEAP activity in NOD2 G908R cells. NOD2 G908R cells were treated for 24 h with 0.2 μg/ml MDP in the presence or absence of L- (A) or D, L-SFN (B). Media was collected and added to QUANTI-Blue™ solution and left at room temperature for 5-15 min before being measured using a spectrophotometer at 650 nm. Data shown = mean ± SD, data representative of three independent experiments. + p < 0.05, ++ p < 0.01, +++ p < 0.001 comparison to 0 μM –MDP. *p < 0.05, **p < 0.01, ***p < 0.001, comparison to 0 μM + MDP.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4335778&req=5

Fig5: L- and D, L-SFN are able to significantly suppress MDP-induced SEAP activity in NOD2 G908R cells. NOD2 G908R cells were treated for 24 h with 0.2 μg/ml MDP in the presence or absence of L- (A) or D, L-SFN (B). Media was collected and added to QUANTI-Blue™ solution and left at room temperature for 5-15 min before being measured using a spectrophotometer at 650 nm. Data shown = mean ± SD, data representative of three independent experiments. + p < 0.05, ++ p < 0.01, +++ p < 0.001 comparison to 0 μM –MDP. *p < 0.05, **p < 0.01, ***p < 0.001, comparison to 0 μM + MDP.
Mentions: We subsequently confirmed if there was a differential effect based on SFN isomer in the NOD2 WT cells (Figure 4). The only significant difference was seen at 10 μM SFN stimulated with 0.2 μg/ml MDP (Figure 4). This was not as a result of reduced viability with D, L-SFN (data not shown). Finally we examined if the same suppression was true in NOD2 G908R cells. Similarly to the NOD2 WT cells, both SFN isomers showed significant reductions at nearly all the concentrations used (Figure 5). No significant differences were found between the suppression of MDP-induced NF-κB activity when using the L- or the D, L-SFN forms (Figure 6).Figure 4

Bottom Line: These results raise the possibility that PRR-mediated inflammation could be suppressed by specific dietary bioactives.The cells were co-treated with sulforaphane and MDP and secreted alkaline phosphatase (SEAP) production was determined.These results demonstrate that the anti-inflammatory role of sulforaphane is not restricted to LPS-induced inflammatory signalling.

View Article: PubMed Central - PubMed

Affiliation: Food and Health Programme, Institute of Food Research, Norwich Research Park, Norwich, UK.

ABSTRACT

Background: Sulforaphane has well established anti-cancer properties and more recently anti-inflammatory properties have also been determined. Sulforaphane has been shown to inhibit PRR-mediated pro-inflammatory signalling by either directly targeting the receptor or their downstream signalling molecules such as the transcription factor, NF-κB. These results raise the possibility that PRR-mediated inflammation could be suppressed by specific dietary bioactives. We examined whether sulforaphane could suppress NF-κB via the NOD2 pathway.

Methods: Human embryonic kidney 293T (HEK293T) cells were stably transfected with NOD2 variants and the NF-κB reporter, pNifty2-SEAP. The cells were co-treated with sulforaphane and MDP and secreted alkaline phosphatase (SEAP) production was determined.

Results: We found that sulforaphane was able to significantly suppress the ligand-induced NF-κB activity at physiologically relevant concentrations, achievable via the consumption of broccoli within the diet.

Conclusions: These results demonstrate that the anti-inflammatory role of sulforaphane is not restricted to LPS-induced inflammatory signalling. These data add to the growing evidence that PRR activation can be inhibited by specific phytochemicals and thus suggests that diet could be a way of controlling inflammation. This is particularly important for a disease like Crohn's disease where diet can play a key role in relieving or exacerbating symptoms.

No MeSH data available.


Related in: MedlinePlus