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Chronic Heavy Alcohol Use is Associated with Upregulated Paneth Cell Antimicrobials in Gastric Mucosa.

Ostaff MJ, Schäfer C, Courth L, Stebe SR, Ott G, Stange EF, Wehkamp J - Clin Transl Gastroenterol (2015)

Bottom Line: We examined the effect of heavy alcohol use on gastric and duodenal Paneth-cell-derived and epithelial antimicrobial peptides (AMPs), cytokines, and the Wnt pathway, an important regulator of epithelial regeneration.No significant differences were found for β-defensins and cytokines (interleukins IL1β, IL6, IL8, IL10).This may be linked to Wnt pathway activation, which has an important role in the epithelial regenerative homeostasis.

View Article: PubMed Central - PubMed

Affiliation: Dr Margarete-Fischer-Bosch-Institute of Clinical Pharmacology, Stuttgart, and University of Tübingen, Tübingen, Germany.

ABSTRACT

Objectives: How alcohol consumption affects the integrity and the defense mechanisms of the mucosa in the upper gastrointestinal tract is largely unknown. We examined the effect of heavy alcohol use on gastric and duodenal Paneth-cell-derived and epithelial antimicrobial peptides (AMPs), cytokines, and the Wnt pathway, an important regulator of epithelial regeneration.

Methods: In 22 patients with heavy alcohol use and 17 control subjects, biopsies from gastric corpus, antrum, and duodenum were examined for messenger RNA (mRNA) of AMPs, cytokines, and Wnt pathway factors using real-time PCR. The expression of the α-defensin HD5 was analyzed immunohistochemically. The effect of alcohol exposure on Wnt signaling and AMP production was also studied in a gastric cell line using mRNA and reporter gene assays.

Results: Heavy alcohol use was associated with increased expression of Paneth cell HD5 and HD6 mRNA in the antrum, where these products are normally absent (HD5 mRNA in controls vs.

Patients: 2100±900 and 365 500±161 600, HD6 mRNA: 320±130 and 58 300±32 600 copies per 10 ng total RNA, means±s.e.m., P value: 0.022 and 0.011). Upregulated HD5 was independent of intestinal metaplasia that was observed in a minority of patients. No significant differences were found for β-defensins and cytokines (interleukins IL1β, IL6, IL8, IL10). In patients, Wnt pathway factors showed a trend toward higher levels. In vitro, ethanol exposure induced the production of HD5 and HD6 and activation of the Wnt pathway.

Conclusions: Alcohol exposure can induce gastric Paneth cell AMP expression. This may be linked to Wnt pathway activation, which has an important role in the epithelial regenerative homeostasis.

No MeSH data available.


Related in: MedlinePlus

Fold induction of reporter gene activity of HD5 (a), HD6 (b), and the Wnt responsive TopFlash promotor (c) in response to a 20 h stimulation with different ethanol concentrations in the culture media. In addition, a regular as well as a mutated HD5 promotor, lacking three essential binding sites for TCF/Lef Wnt transcription factors, were stimulated 20 h by different concentrations of H2O2, with and without a parallel treatment of 5% EtOH (d). Luciferase reporter experiments were carried out in triplicates. Shown are means of independent experiments with the respective s.e.m. Significances: Friedman test P<0.0001 for (a), (b), and (d), P=0.006 for (c), Dunn's post test: *P<0.05, **P<0.01, ***P<0.001.
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fig7: Fold induction of reporter gene activity of HD5 (a), HD6 (b), and the Wnt responsive TopFlash promotor (c) in response to a 20 h stimulation with different ethanol concentrations in the culture media. In addition, a regular as well as a mutated HD5 promotor, lacking three essential binding sites for TCF/Lef Wnt transcription factors, were stimulated 20 h by different concentrations of H2O2, with and without a parallel treatment of 5% EtOH (d). Luciferase reporter experiments were carried out in triplicates. Shown are means of independent experiments with the respective s.e.m. Significances: Friedman test P<0.0001 for (a), (b), and (d), P=0.006 for (c), Dunn's post test: *P<0.05, **P<0.01, ***P<0.001.

Mentions: Graphs that are presented as means±s.e.m. were generated using GraphPad Prism V6 software. Differences in patient characteristics and in the mRNA analyses were analyzed for statistical significance by Mann–Whitney U-test (for non-normally distributed groups). To compare percentages or absolute cases, Fisher's exact test was performed. To adjust for multiple comparisons we used the Bonferroni correction. To test for significance of the luciferase reporter gene fold inductions, we used a matched, non-parametric one-way analysis (Friedman Test) and performed post analysis via Dunn's test for the induction by 1 and 5% EtOH (Figure 7a) and for the induction of HD5 and HD5mut by 5% EtOH under all H2O2 concentrations (Figure 7d). P values below 0.05 were considered statistically significant (P<0.05=* P<0.01=**, P<0.001=***).


Chronic Heavy Alcohol Use is Associated with Upregulated Paneth Cell Antimicrobials in Gastric Mucosa.

Ostaff MJ, Schäfer C, Courth L, Stebe SR, Ott G, Stange EF, Wehkamp J - Clin Transl Gastroenterol (2015)

Fold induction of reporter gene activity of HD5 (a), HD6 (b), and the Wnt responsive TopFlash promotor (c) in response to a 20 h stimulation with different ethanol concentrations in the culture media. In addition, a regular as well as a mutated HD5 promotor, lacking three essential binding sites for TCF/Lef Wnt transcription factors, were stimulated 20 h by different concentrations of H2O2, with and without a parallel treatment of 5% EtOH (d). Luciferase reporter experiments were carried out in triplicates. Shown are means of independent experiments with the respective s.e.m. Significances: Friedman test P<0.0001 for (a), (b), and (d), P=0.006 for (c), Dunn's post test: *P<0.05, **P<0.01, ***P<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4816256&req=5

fig7: Fold induction of reporter gene activity of HD5 (a), HD6 (b), and the Wnt responsive TopFlash promotor (c) in response to a 20 h stimulation with different ethanol concentrations in the culture media. In addition, a regular as well as a mutated HD5 promotor, lacking three essential binding sites for TCF/Lef Wnt transcription factors, were stimulated 20 h by different concentrations of H2O2, with and without a parallel treatment of 5% EtOH (d). Luciferase reporter experiments were carried out in triplicates. Shown are means of independent experiments with the respective s.e.m. Significances: Friedman test P<0.0001 for (a), (b), and (d), P=0.006 for (c), Dunn's post test: *P<0.05, **P<0.01, ***P<0.001.
Mentions: Graphs that are presented as means±s.e.m. were generated using GraphPad Prism V6 software. Differences in patient characteristics and in the mRNA analyses were analyzed for statistical significance by Mann–Whitney U-test (for non-normally distributed groups). To compare percentages or absolute cases, Fisher's exact test was performed. To adjust for multiple comparisons we used the Bonferroni correction. To test for significance of the luciferase reporter gene fold inductions, we used a matched, non-parametric one-way analysis (Friedman Test) and performed post analysis via Dunn's test for the induction by 1 and 5% EtOH (Figure 7a) and for the induction of HD5 and HD5mut by 5% EtOH under all H2O2 concentrations (Figure 7d). P values below 0.05 were considered statistically significant (P<0.05=* P<0.01=**, P<0.001=***).

Bottom Line: We examined the effect of heavy alcohol use on gastric and duodenal Paneth-cell-derived and epithelial antimicrobial peptides (AMPs), cytokines, and the Wnt pathway, an important regulator of epithelial regeneration.No significant differences were found for β-defensins and cytokines (interleukins IL1β, IL6, IL8, IL10).This may be linked to Wnt pathway activation, which has an important role in the epithelial regenerative homeostasis.

View Article: PubMed Central - PubMed

Affiliation: Dr Margarete-Fischer-Bosch-Institute of Clinical Pharmacology, Stuttgart, and University of Tübingen, Tübingen, Germany.

ABSTRACT

Objectives: How alcohol consumption affects the integrity and the defense mechanisms of the mucosa in the upper gastrointestinal tract is largely unknown. We examined the effect of heavy alcohol use on gastric and duodenal Paneth-cell-derived and epithelial antimicrobial peptides (AMPs), cytokines, and the Wnt pathway, an important regulator of epithelial regeneration.

Methods: In 22 patients with heavy alcohol use and 17 control subjects, biopsies from gastric corpus, antrum, and duodenum were examined for messenger RNA (mRNA) of AMPs, cytokines, and Wnt pathway factors using real-time PCR. The expression of the α-defensin HD5 was analyzed immunohistochemically. The effect of alcohol exposure on Wnt signaling and AMP production was also studied in a gastric cell line using mRNA and reporter gene assays.

Results: Heavy alcohol use was associated with increased expression of Paneth cell HD5 and HD6 mRNA in the antrum, where these products are normally absent (HD5 mRNA in controls vs.

Patients: 2100±900 and 365 500±161 600, HD6 mRNA: 320±130 and 58 300±32 600 copies per 10 ng total RNA, means±s.e.m., P value: 0.022 and 0.011). Upregulated HD5 was independent of intestinal metaplasia that was observed in a minority of patients. No significant differences were found for β-defensins and cytokines (interleukins IL1β, IL6, IL8, IL10). In patients, Wnt pathway factors showed a trend toward higher levels. In vitro, ethanol exposure induced the production of HD5 and HD6 and activation of the Wnt pathway.

Conclusions: Alcohol exposure can induce gastric Paneth cell AMP expression. This may be linked to Wnt pathway activation, which has an important role in the epithelial regenerative homeostasis.

No MeSH data available.


Related in: MedlinePlus