Limits...
Sulforaphane epigenetically regulates innate immune responses of porcine monocyte-derived dendritic cells induced with lipopolysaccharide.

Qu X, Pröll M, Neuhoff C, Zhang R, Cinar MU, Hossain MM, Tesfaye D, Große-Brinkhaus C, Salilew-Wondim D, Tholen E, Looft C, Hölker M, Schellander K, Uddin MJ - PLoS ONE (2015)

Bottom Line: Although dendritic cells (DCs) are playing pivotal roles in host immune responses, the effect of epigenetic modulation of DCs immune responses remains unknown.SFN was found to inhibit the lipopolysaccharide LPS induced HDAC6, HDAC10 and DNA methyltransferase (DNMT3a) gene expression, whereas up-regulated the expression of DNMT1 gene.SFN impaired the pro-inflammatory cytokine TNF-α and IL-1ß secretion into the cell culture supernatants that were induced in moDCs by LPS stimulation, whereas SFN increased the cellular-resident TNF-α accumulation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115 Bonn, Germany.

ABSTRACT
Histone acetylation, regulated by histone deacetylases (HDACs) is a key epigenetic mechanism controlling gene expressions. Although dendritic cells (DCs) are playing pivotal roles in host immune responses, the effect of epigenetic modulation of DCs immune responses remains unknown. Sulforaphane (SFN) as a HDAC inhibitor has anti-inflammatory properties, which is used to investigate the epigenetic regulation of LPS-induced immune gene and HDAC family gene expressions in porcine monocyte-derived dendritic cells (moDCs). SFN was found to inhibit the lipopolysaccharide LPS induced HDAC6, HDAC10 and DNA methyltransferase (DNMT3a) gene expression, whereas up-regulated the expression of DNMT1 gene. Additionally, SFN was observed to inhibit the global HDAC activity, and suppressed moDCs differentiation from immature to mature DCs through down-regulating the CD40, CD80 and CD86 expression and led further to enhanced phagocytosis of moDCs. The SFN pre-treated of moDCs directly altered the LPS-induced TLR4 and MD2 gene expression and dynamically regulated the TLR4-induced activity of transcription factor NF-κB and TBP. SFN showed a protective role in LPS induced cell apoptosis through suppressing the IRF6 and TGF-ß1 production. SFN impaired the pro-inflammatory cytokine TNF-α and IL-1ß secretion into the cell culture supernatants that were induced in moDCs by LPS stimulation, whereas SFN increased the cellular-resident TNF-α accumulation. This study demonstrates that through the epigenetic mechanism the HDAC inhibitor SFN could modulate the LPS induced innate immune responses of porcine moDCs.

No MeSH data available.


Related in: MedlinePlus

SFN inhibits HDAC activity and regulates genes which encode epigenetic enzymes.moDCs at day 7 in cell culture were used for this experiment. Relative HDAC activity assay was measured using the Color-de-Lys HDAC colorimetric activity assay kit. To confirm the global HDAC deacetylation of moDCs, cells were stimulated with different concentration of SFN (Control, 5 μM, 10 μM, 15 μM, and 20 μM) for 24 h (A). Equal amounts of isolated nuclear protein were subjected to HDAC activity analysis. The effects of SFN (10 μM) on gene expression of epigenetic encoding enzymes in porcine moDCs stimulated with LPS were examined. The HDAC6 (B), HDAC10 (C), DNMT3a (D) and DNMT1 (E) mRNA expression was quantified using qRT-PCR. The moDCs were pre-treated for 24 h with or without SFN before stimulating with LPS (1 μg/ml) for additional 24 h. The results (A, B, C, D, and E) were represented as the mean ± standard deviation (SD) of three independent experiments and each experiment was performed in duplicate (*p < 0.05; **p < 0.01; ***p < 0.001).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4368608&req=5

pone.0121574.g004: SFN inhibits HDAC activity and regulates genes which encode epigenetic enzymes.moDCs at day 7 in cell culture were used for this experiment. Relative HDAC activity assay was measured using the Color-de-Lys HDAC colorimetric activity assay kit. To confirm the global HDAC deacetylation of moDCs, cells were stimulated with different concentration of SFN (Control, 5 μM, 10 μM, 15 μM, and 20 μM) for 24 h (A). Equal amounts of isolated nuclear protein were subjected to HDAC activity analysis. The effects of SFN (10 μM) on gene expression of epigenetic encoding enzymes in porcine moDCs stimulated with LPS were examined. The HDAC6 (B), HDAC10 (C), DNMT3a (D) and DNMT1 (E) mRNA expression was quantified using qRT-PCR. The moDCs were pre-treated for 24 h with or without SFN before stimulating with LPS (1 μg/ml) for additional 24 h. The results (A, B, C, D, and E) were represented as the mean ± standard deviation (SD) of three independent experiments and each experiment was performed in duplicate (*p < 0.05; **p < 0.01; ***p < 0.001).

Mentions: We investigated the HDAC activity in different concentration of SFN treatments. The results showed that SFN significantly inhibited HDAC activity in a dose-dependent manner (Fig. 4A). The effects of SFN on genes encoding epigenetic enzyme showed that SFN treatment caused a decrease in mRNA expression of several HDAC genes in LPS treated porcine moDCs (Fig. 4B and C). Our results show that SFN significantly inhibited both HDAC6 and HDAC10 mRNA expression that were induced by LPS in moDCs (Fig. 4B and C). Similarly, SFN treatment significantly enhanced the down-regulation of de novo methyltransferase DNMT3a that was induced by LPS treatment (Fig. 4D). The DNMT1 expression was significantly increased in SFN pre-treated moDCs that was induced by LPS treatment (Fig. 4E). However, this trend could not be observed in the case of other (data not shown) DNMT genes.


Sulforaphane epigenetically regulates innate immune responses of porcine monocyte-derived dendritic cells induced with lipopolysaccharide.

Qu X, Pröll M, Neuhoff C, Zhang R, Cinar MU, Hossain MM, Tesfaye D, Große-Brinkhaus C, Salilew-Wondim D, Tholen E, Looft C, Hölker M, Schellander K, Uddin MJ - PLoS ONE (2015)

SFN inhibits HDAC activity and regulates genes which encode epigenetic enzymes.moDCs at day 7 in cell culture were used for this experiment. Relative HDAC activity assay was measured using the Color-de-Lys HDAC colorimetric activity assay kit. To confirm the global HDAC deacetylation of moDCs, cells were stimulated with different concentration of SFN (Control, 5 μM, 10 μM, 15 μM, and 20 μM) for 24 h (A). Equal amounts of isolated nuclear protein were subjected to HDAC activity analysis. The effects of SFN (10 μM) on gene expression of epigenetic encoding enzymes in porcine moDCs stimulated with LPS were examined. The HDAC6 (B), HDAC10 (C), DNMT3a (D) and DNMT1 (E) mRNA expression was quantified using qRT-PCR. The moDCs were pre-treated for 24 h with or without SFN before stimulating with LPS (1 μg/ml) for additional 24 h. The results (A, B, C, D, and E) were represented as the mean ± standard deviation (SD) of three independent experiments and each experiment was performed in duplicate (*p < 0.05; **p < 0.01; ***p < 0.001).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368608&req=5

pone.0121574.g004: SFN inhibits HDAC activity and regulates genes which encode epigenetic enzymes.moDCs at day 7 in cell culture were used for this experiment. Relative HDAC activity assay was measured using the Color-de-Lys HDAC colorimetric activity assay kit. To confirm the global HDAC deacetylation of moDCs, cells were stimulated with different concentration of SFN (Control, 5 μM, 10 μM, 15 μM, and 20 μM) for 24 h (A). Equal amounts of isolated nuclear protein were subjected to HDAC activity analysis. The effects of SFN (10 μM) on gene expression of epigenetic encoding enzymes in porcine moDCs stimulated with LPS were examined. The HDAC6 (B), HDAC10 (C), DNMT3a (D) and DNMT1 (E) mRNA expression was quantified using qRT-PCR. The moDCs were pre-treated for 24 h with or without SFN before stimulating with LPS (1 μg/ml) for additional 24 h. The results (A, B, C, D, and E) were represented as the mean ± standard deviation (SD) of three independent experiments and each experiment was performed in duplicate (*p < 0.05; **p < 0.01; ***p < 0.001).
Mentions: We investigated the HDAC activity in different concentration of SFN treatments. The results showed that SFN significantly inhibited HDAC activity in a dose-dependent manner (Fig. 4A). The effects of SFN on genes encoding epigenetic enzyme showed that SFN treatment caused a decrease in mRNA expression of several HDAC genes in LPS treated porcine moDCs (Fig. 4B and C). Our results show that SFN significantly inhibited both HDAC6 and HDAC10 mRNA expression that were induced by LPS in moDCs (Fig. 4B and C). Similarly, SFN treatment significantly enhanced the down-regulation of de novo methyltransferase DNMT3a that was induced by LPS treatment (Fig. 4D). The DNMT1 expression was significantly increased in SFN pre-treated moDCs that was induced by LPS treatment (Fig. 4E). However, this trend could not be observed in the case of other (data not shown) DNMT genes.

Bottom Line: Although dendritic cells (DCs) are playing pivotal roles in host immune responses, the effect of epigenetic modulation of DCs immune responses remains unknown.SFN was found to inhibit the lipopolysaccharide LPS induced HDAC6, HDAC10 and DNA methyltransferase (DNMT3a) gene expression, whereas up-regulated the expression of DNMT1 gene.SFN impaired the pro-inflammatory cytokine TNF-α and IL-1ß secretion into the cell culture supernatants that were induced in moDCs by LPS stimulation, whereas SFN increased the cellular-resident TNF-α accumulation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115 Bonn, Germany.

ABSTRACT
Histone acetylation, regulated by histone deacetylases (HDACs) is a key epigenetic mechanism controlling gene expressions. Although dendritic cells (DCs) are playing pivotal roles in host immune responses, the effect of epigenetic modulation of DCs immune responses remains unknown. Sulforaphane (SFN) as a HDAC inhibitor has anti-inflammatory properties, which is used to investigate the epigenetic regulation of LPS-induced immune gene and HDAC family gene expressions in porcine monocyte-derived dendritic cells (moDCs). SFN was found to inhibit the lipopolysaccharide LPS induced HDAC6, HDAC10 and DNA methyltransferase (DNMT3a) gene expression, whereas up-regulated the expression of DNMT1 gene. Additionally, SFN was observed to inhibit the global HDAC activity, and suppressed moDCs differentiation from immature to mature DCs through down-regulating the CD40, CD80 and CD86 expression and led further to enhanced phagocytosis of moDCs. The SFN pre-treated of moDCs directly altered the LPS-induced TLR4 and MD2 gene expression and dynamically regulated the TLR4-induced activity of transcription factor NF-κB and TBP. SFN showed a protective role in LPS induced cell apoptosis through suppressing the IRF6 and TGF-ß1 production. SFN impaired the pro-inflammatory cytokine TNF-α and IL-1ß secretion into the cell culture supernatants that were induced in moDCs by LPS stimulation, whereas SFN increased the cellular-resident TNF-α accumulation. This study demonstrates that through the epigenetic mechanism the HDAC inhibitor SFN could modulate the LPS induced innate immune responses of porcine moDCs.

No MeSH data available.


Related in: MedlinePlus