Limits...
Microbial metabolite butyrate facilitates M2 macrophage polarization and function.

Ji J, Shu D, Zheng M, Wang J, Luo C, Wang Y, Guo F, Zou X, Lv X, Li Y, Liu T, Qu H - Sci Rep (2016)

Bottom Line: Metabolites from intestinal microbes modulate the mucosal immune system by regulating the polarization and expansion of T cells.Butyrate attenuated intestinal inflammation in mice with dextran sulfate sodium (DSS)-induced colitis, with a significant increase in colonic expression of the M2 macrophage-associated protein, Arg1.Collectively, our study indicated that commensal microbe-derived butyrate is a novel activator of STAT6-mediated transcription through H3K9 acetylation driving M2 macrophage polarization, and delineated new insights into the immune interplay underlying inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Institute of Animal Science, Guangdong Academy of Agricultural Sciences, 1 Dafeng 1st Street, Wushan, Tianhe District, Guangzhou 510640, China.

ABSTRACT
Metabolites from intestinal microbes modulate the mucosal immune system by regulating the polarization and expansion of T cells. Whether the microbial metabolites influence macrophage polarization, however, is poorly understood. Here, we show that the large bowel microbial fermentation product, butyrate, facilitates M2 macrophage polarization, in vitro and in vivo. The supernatant from butyrate-treated M2 macrophage increased the migration and enhanced the wound closure rate of MLE-12 cells. Butyrate attenuated intestinal inflammation in mice with dextran sulfate sodium (DSS)-induced colitis, with a significant increase in colonic expression of the M2 macrophage-associated protein, Arg1. M2 macrophage treated with butyrate, had increased activation of the H3K9/STAT6 signaling pathway, suggesting a mechanism for butyrate facilitated M2 macrophage polarization. Collectively, our study indicated that commensal microbe-derived butyrate is a novel activator of STAT6-mediated transcription through H3K9 acetylation driving M2 macrophage polarization, and delineated new insights into the immune interplay underlying inflammatory bowel disease.

No MeSH data available.


Related in: MedlinePlus

Histone acetyltransferase (HAT) inhibition mediates butyrate-treated M2 macrophage polarization by inhibiting the expression of acetylated H3K9.(a) Arg1 and Ym1 gene expression in butyrate-treated M2 macrophage treated with increasing concentrations (2–20 μM) of HAT inhibitor, C646. (b) Arg1 and Ym1 gene expression in the presence of butyrate (50 μg/ml) and C646 (5 μM). (c) Western blot analysis of Arg1 protein in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). (d) Quantitative RT-PCR analysis of HDAC1 gene expression in butyrate-treated M2-BMDMs after 6 h treatment with C646 (5 μM). (e) Western blot analysis of acetylated H3K9, total histone H3, and β-actin proteins in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). Cells were lysed and western blotting performed with the indicated antibodies. (f) Phosphorylation of STAT6 in butyrate-treated M2-BMDMs. Western blotting was performed with anti-phospho-STAT6, STAT6, and β-actin. Data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4837405&req=5

f7: Histone acetyltransferase (HAT) inhibition mediates butyrate-treated M2 macrophage polarization by inhibiting the expression of acetylated H3K9.(a) Arg1 and Ym1 gene expression in butyrate-treated M2 macrophage treated with increasing concentrations (2–20 μM) of HAT inhibitor, C646. (b) Arg1 and Ym1 gene expression in the presence of butyrate (50 μg/ml) and C646 (5 μM). (c) Western blot analysis of Arg1 protein in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). (d) Quantitative RT-PCR analysis of HDAC1 gene expression in butyrate-treated M2-BMDMs after 6 h treatment with C646 (5 μM). (e) Western blot analysis of acetylated H3K9, total histone H3, and β-actin proteins in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). Cells were lysed and western blotting performed with the indicated antibodies. (f) Phosphorylation of STAT6 in butyrate-treated M2-BMDMs. Western blotting was performed with anti-phospho-STAT6, STAT6, and β-actin. Data are representative of three independent experiments.

Mentions: To investigate whether acetylation of H3K9 was involved in M2 macrophage polarization, the p300/CBP specific histone acetyltransferase (HAT) inhibitor, C646, was used to reduce the acetylation of H3K9. We observed that with increasing concentrations (2–20 μM) of C646, Arg1 and Ym1 mRNA expression was decreased (Fig. 7a). Additionally, C646 inhibited the polarization of M2-BMDMs and M2-BMDMs treated with butyrate (Fig. 7b). Additionally, the Arg1 protein level was dramatically decreased in butyrate-treated M2-BMDMs with concomitant C646 treatment, compared with untreated M2-BMDM controls (Fig. 7c). Meanwhile, C646 effectively enhanced the expression of HDAC1 and reduced the acetylation of H3K9 in both M2-BMDMs and butyrate-treated M2-BMDMs (Fig. 7d,e). Moreover, we found lower STAT6 phosphorylation in response to C646 in butyrate-treated M2-BMDMs compared with untreated M2-BMDMs. Together, we concluded that butyrate mediated its effects on M2 macrophage largely through its HDAC inhibitor activity augmenting M2 macrophage polarization by the STAT6 signaling pathway.


Microbial metabolite butyrate facilitates M2 macrophage polarization and function.

Ji J, Shu D, Zheng M, Wang J, Luo C, Wang Y, Guo F, Zou X, Lv X, Li Y, Liu T, Qu H - Sci Rep (2016)

Histone acetyltransferase (HAT) inhibition mediates butyrate-treated M2 macrophage polarization by inhibiting the expression of acetylated H3K9.(a) Arg1 and Ym1 gene expression in butyrate-treated M2 macrophage treated with increasing concentrations (2–20 μM) of HAT inhibitor, C646. (b) Arg1 and Ym1 gene expression in the presence of butyrate (50 μg/ml) and C646 (5 μM). (c) Western blot analysis of Arg1 protein in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). (d) Quantitative RT-PCR analysis of HDAC1 gene expression in butyrate-treated M2-BMDMs after 6 h treatment with C646 (5 μM). (e) Western blot analysis of acetylated H3K9, total histone H3, and β-actin proteins in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). Cells were lysed and western blotting performed with the indicated antibodies. (f) Phosphorylation of STAT6 in butyrate-treated M2-BMDMs. Western blotting was performed with anti-phospho-STAT6, STAT6, and β-actin. Data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837405&req=5

f7: Histone acetyltransferase (HAT) inhibition mediates butyrate-treated M2 macrophage polarization by inhibiting the expression of acetylated H3K9.(a) Arg1 and Ym1 gene expression in butyrate-treated M2 macrophage treated with increasing concentrations (2–20 μM) of HAT inhibitor, C646. (b) Arg1 and Ym1 gene expression in the presence of butyrate (50 μg/ml) and C646 (5 μM). (c) Western blot analysis of Arg1 protein in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). (d) Quantitative RT-PCR analysis of HDAC1 gene expression in butyrate-treated M2-BMDMs after 6 h treatment with C646 (5 μM). (e) Western blot analysis of acetylated H3K9, total histone H3, and β-actin proteins in butyrate-treated M2-BMDMs after 12 h treatment with C646 (5 μM). Cells were lysed and western blotting performed with the indicated antibodies. (f) Phosphorylation of STAT6 in butyrate-treated M2-BMDMs. Western blotting was performed with anti-phospho-STAT6, STAT6, and β-actin. Data are representative of three independent experiments.
Mentions: To investigate whether acetylation of H3K9 was involved in M2 macrophage polarization, the p300/CBP specific histone acetyltransferase (HAT) inhibitor, C646, was used to reduce the acetylation of H3K9. We observed that with increasing concentrations (2–20 μM) of C646, Arg1 and Ym1 mRNA expression was decreased (Fig. 7a). Additionally, C646 inhibited the polarization of M2-BMDMs and M2-BMDMs treated with butyrate (Fig. 7b). Additionally, the Arg1 protein level was dramatically decreased in butyrate-treated M2-BMDMs with concomitant C646 treatment, compared with untreated M2-BMDM controls (Fig. 7c). Meanwhile, C646 effectively enhanced the expression of HDAC1 and reduced the acetylation of H3K9 in both M2-BMDMs and butyrate-treated M2-BMDMs (Fig. 7d,e). Moreover, we found lower STAT6 phosphorylation in response to C646 in butyrate-treated M2-BMDMs compared with untreated M2-BMDMs. Together, we concluded that butyrate mediated its effects on M2 macrophage largely through its HDAC inhibitor activity augmenting M2 macrophage polarization by the STAT6 signaling pathway.

Bottom Line: Metabolites from intestinal microbes modulate the mucosal immune system by regulating the polarization and expansion of T cells.Butyrate attenuated intestinal inflammation in mice with dextran sulfate sodium (DSS)-induced colitis, with a significant increase in colonic expression of the M2 macrophage-associated protein, Arg1.Collectively, our study indicated that commensal microbe-derived butyrate is a novel activator of STAT6-mediated transcription through H3K9 acetylation driving M2 macrophage polarization, and delineated new insights into the immune interplay underlying inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Institute of Animal Science, Guangdong Academy of Agricultural Sciences, 1 Dafeng 1st Street, Wushan, Tianhe District, Guangzhou 510640, China.

ABSTRACT
Metabolites from intestinal microbes modulate the mucosal immune system by regulating the polarization and expansion of T cells. Whether the microbial metabolites influence macrophage polarization, however, is poorly understood. Here, we show that the large bowel microbial fermentation product, butyrate, facilitates M2 macrophage polarization, in vitro and in vivo. The supernatant from butyrate-treated M2 macrophage increased the migration and enhanced the wound closure rate of MLE-12 cells. Butyrate attenuated intestinal inflammation in mice with dextran sulfate sodium (DSS)-induced colitis, with a significant increase in colonic expression of the M2 macrophage-associated protein, Arg1. M2 macrophage treated with butyrate, had increased activation of the H3K9/STAT6 signaling pathway, suggesting a mechanism for butyrate facilitated M2 macrophage polarization. Collectively, our study indicated that commensal microbe-derived butyrate is a novel activator of STAT6-mediated transcription through H3K9 acetylation driving M2 macrophage polarization, and delineated new insights into the immune interplay underlying inflammatory bowel disease.

No MeSH data available.


Related in: MedlinePlus