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. 2019 Dec;176(23):4446-4461.
doi: 10.1111/bph.14806. Epub 2019 Dec 8.

Butyrate ameliorates caerulein-induced acute pancreatitis and associated intestinal injury by tissue-specific mechanisms

Affiliations

Butyrate ameliorates caerulein-induced acute pancreatitis and associated intestinal injury by tissue-specific mechanisms

Xiaohua Pan et al. Br J Pharmacol. 2019 Dec.

Abstract

Background and purpose: Acute pancreatitis (AP) is a common acute abdominal condition, frequently associated with intestinal barrier dysfunction, which aggravates AP retroactively. Butyrate exhibits anti-inflammatory effects in a variety of inflammatory diseases. However, its potential beneficial effect on AP and the underlying mechanisms have not been investigated.

Experimental approach: Experimental AP was induced by caerulein hyperstimulation in wild-type and GPR109A-/- mice. Sodium butyrate was administered intragastrically for 7 days prior to caerulein hyperstimulation. Anti-inflammatory mechanisms of butyrate were further investigated in peritoneal macrophages.

Key results: Butyrate prophylaxis attenuated AP as shown by reduced serum amylase and lipase levels, pancreatic oedema, myeloperoxidase activity, and improved pancreatic morphology. Amelioration of pancreatic damage by butyrate was associated with reduced levels of TNF-α, IL-6, and CCL2 and suppressed activation of the NLRP3 inflammasome in both pancreas and colon. Further, butyrate ameliorated pancreatic inflammation by suppressing interactions between histone deacetylase 1 (HDAC1) and AP1 and STAT1 with increased histone acetylation at H3K9, H3K14, H3K18, and H3K27 loci, resulting in suppression of NLRP3 inflammasome activation and modulation of immune cell infiltration in pancreas. Additionally, butyrate mediated STAT1/AP1-NLRP3 inflammasome suppression via HDAC1 inhibition was demonstrated in peritoneal macrophage. In colon, butyrate inhibited NLRP3 inflammasome activation via GPR109A. Accordingly, the modulatory effects of butyrate on AP, AP-associated gut dysfunction, and NLRP3 inflammasome activation were diminished in GPR109A-/- mice.

Conclusion and implications: Our study dissected tissue-specific anti-inflammatory mechanisms of butyrate during AP, suggesting that increased colonic levels of butyrate may be a strategy to protect against AP.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Prophylactic treatment with oral butyrate mitigates the severity of AP. Mice were challenged with PBS (CON), caerulein (CAE), or 50 and 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). Levels of serum amylase (a) and lipase (b), pancreatic oedema (c), and myeloperoxidase (MPO) activity (d) in pancreas. (e) Representative histopathological sections of pancreatic tissues by haematoxylin–eosin staining. Scale bar: 50 μm. (f) ELISA analyses of TNF‐α, IL‐6, and CCL2 levels in pancreas. (g) GC–MS detection of butyrate levels in colon, serum, and pancreas. Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; one‐way ANOVA followed by Tukey's post hoc test
Figure 2
Figure 2
Butyrate prophylaxis decreases innate immune cell infiltration into pancreas during AP. Mice were challenged with PBS (CON), caerulein (CAE), or 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). The frequency of (a) total macrophages (CD45+CD11b+F4/80+), (b) M2 macrophages (CD45+CD11b+F4/80+CD206+), and (c) neutrophils (CD45+CD11b+Ly6G+) in pancreas was detected by flow cytometry. Data shown are means ± SEM, n = 6. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; one‐way ANOVA followed by Tukey's post hoc test
Figure 3
Figure 3
Butyrate attenuates AP‐associated colon injury. Mice were challenged with PBS (CON), caerulein (CAE), or 50 and 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). (a) Representative histopathological sections of colon by haematoxylin–eosin staining. Scale bar: 100 μm. (b) Colonic crypt length. (c) ELISA detection of TNF‐α, IL‐6, and CCL2 levels in colon. (d) Serum diamine oxidase (DAO) activity. Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; one‐way ANOVA followed by Tukey's post hoc test
Figure 4
Figure 4
Butyrate attenuates pancreatic and colonic injury and inflammatory response by modulating NLRP3 inflammasome pathway. Mice were challenged with PBS (CON), caerulein (CAE), or 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). Representative bands of Western blot and quantitative analyses of NLRP3 inflammasome pathway in pancreas (a) and colon (b). Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE
Figure 5
Figure 5
Butyrate blocks the activation of transcription factors. Mice were challenged with PBS (CON), caerulein (CAE), or 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). Representative bands of Western blot and phosphorylation quantification of transcription factors NF‐κB, AP1, STAT1, and STAT3 in pancreas (a) and colons (b). Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; one‐way ANOVA followed by Tukey's post hoc test
Figure 6
Figure 6
Butyrate exerts anti‐inflammatory effects by acting as HDAC inhibitor in the pancreas but not in colon. Mice were challenged with PBS (CON), caerulein (CAE), or 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). Protein expression of HDAC1, 2, and 3 in pancreas (a) and colon (c) was determined by Western blot. Histone acetylation at H3K9, H3K14, H3K18, H3K27, and H3K56 in pancreas (b) and colon (d) was detected by Western blot. Co‐IP analysis of HDAC1 with NF‐κB (p65), AP1, STAT1, and STAT3 in pancreas (e) and colon (f). Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; one‐way ANOVA followed by Tukey's post hoc test
Figure 7
Figure 7
Butyrate inhibits AP1/STAT1 phosphorylation and NLRP3 inflammasome activation by acting as an HDAC1 inhibitor in peritoneal macrophage. Cells were pretreated with sodium butyrate (SB; 5 μM, 100 μM, and 2 mM) for 24 hr followed by incubation with 100 ng·ml−1 LPS for 6 hr. (a) Representative bands of Western blot. (b) Relative quantitative analyses of HDAC1 expression in macrophages. (c, d) Phosphorylation analyses of transcription factors AP1 (c) and STAT1 (d) in macrophages. (e) Western blotting analysis of NLRP3 inflammasome and associated proteins' expression in macrophages. Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from LPS; one‐way ANOVA followed by Tukey's post hoc test
Figure 8
Figure 8
Butyrate protects against colonic inflammation via its receptors GPR109A, contributing to AP attenuation. Mice were challenged with PBS (CON), caerulein (CAE), or 200 mg·kg−1 sodium butyrate combined with caerulein (SB + CAE). Western blot analysis of GPR109A and GPR41 in pancreas (a) and colon (b). Levels of serum amylase (c) and lipase (d), pancreatic oedema (e), and myeloperoxidase (MPO) activity (f) in pancreas. (g) Serum diamine oxidase (DAO) activity. Western blot analysis of NLRP3 inflammasome and associated protein expression in pancreas (h) and colon (i), and values represented fold change after normalization to control. Data shown are means ± SEM, n = 8. # P < .05, significantly different from CON; *P < .05, significantly different from CAE; $ P < .05, significantly different from CAE; & P < .05, significantly different from SB + CAE; one‐way ANOVA followed by Tukey's post hoc test

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References

    1. Alexander, S. P. H. , Christopoulos, A. , Davenport, A. P. , Kelly, E. , Marrion, N. V. , Peters, J. A. , … CGTP Collaborators (2017). The Concise Guide to PHARMACOLOGY 2017/18: G protein‐coupled receptors. British Journal of Pharmacology, 174, S17–S129. 10.1111/bph.13878 - DOI - PMC - PubMed
    1. Alexander, S. P. H. , Fabbro, D. , Kelly, E. , Marrion, N. V. , Peters, J. A. , Faccenda, E. , … CGTP Collaborators (2017). The Concise Guide to PHARMACOLOGY 2017/18: Enzymes. British Journal of Pharmacology, 174, S272–S359. 10.1111/bph.13877 - DOI - PMC - PubMed
    1. Alexander, S. P. H. , Kelly, E. , Marrion, N. V. , Peters, J. A. , Faccenda, E. , Harding, S. D. , … CGTP Collaborators (2017). The Concise Guide to PHARMACOLOGY 2017/18: Other proteins. British Journal of Pharmacology, 174, S1–S16. 10.1111/bph.13882 - DOI - PMC - PubMed
    1. Alexander, S. P. H. , Roberts, R. E. , Broughton, B. R. S. , Sobey, C. G. , George, C. H. , Stanford, S. C. , … Ahluwalia, A. (2018). Goals and practicalities of immunoblotting and immunohistochemistry: A guide for submission to the British Journal of Pharmacology. British Journal of Pharmacology, 175(3), 407–411. 10.1111/bph.14112 - DOI - PMC - PubMed
    1. Arpaia, N. , Campbell, C. , Fan, X. , Dikiy, S. , van der Veeken, J. , deRoos, P. , … Rudensky, A. Y. (2013). Metabolites produced by commensal bacteria promote peripheral regulatory T‐cell generation. Nature, 504(7480), 451–455. 10.1038/nature12726 - DOI - PMC - PubMed

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