Autophagy Genes of Host Responds to Disruption of Gut Microbial Community by Antibiotics

Dig Dis Sci. 2017 Jun;62(6):1486-1497. doi: 10.1007/s10620-017-4589-8. Epub 2017 May 2.

Abstract

Background: Defective autophagic machinery, such as that in Crohn's disease patients homozygous for ATG16L1 risk allele, is associated with alteration of resident gut bacterial communities. However, whether or not host autophagy responds to changes in the resident gut microbial community is not known. Here, we investigated the effect of antibiotic-induced disruption of the gut microbiome (dysbiosis) on autophagy gene expression and the expression of antimicrobial peptides/protein (AMP) over time.

Aim: To test the hypothesis that antibiotic treatment may cause time-dependent changes in gut bacterial density, autophagy genes, and antimicrobial protein/peptide gene expression.

Methods: Mice (n = 8 per group) were treated with antibiotic cocktail and sacrificed at different intervals of recovery (days 3, 7, 10, 14, 21, 28, 35, and 42) post-antibiotics. DNA and RNA were extracted from small intestinal tissues. Bacterial density, expression of host autophagy genes, and AMP genes were analyzed by relative quantitative PCR. Fold change difference in comparison with untreated control group was calculated using 2-ΔΔCt method. Statistical analysis was performed using nonparametric Mann-Whitney test.

Results: Gut bacterial density changed in a time-dependent fashion in response to antibiotic treatment. These changes were concurrent with upregulation of autophagy genes and antimicrobial peptide/protein gene expression. We further showed that an oral gavage of a resident microbe Desulfovibrio, which bloomed in antibiotic-treated animals, induced Atg5 and lysozyme (Lyz) gene expression.

Conclusion: Autophagy genes respond to dysbiosis induced by antibiotics. This response may be a host mechanism to detect and possibly correct dysbiosis by activating antimicrobial peptides/proteins that control the microbial load in the gut.

Keywords: Antibiotics; Antimicrobial peptide/protein; Autophagy; Desulfovibrio.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Antimicrobial Cationic Peptides / genetics*
  • Autophagy / genetics*
  • Autophagy-Related Protein 5 / genetics
  • Autophagy-Related Proteins / genetics
  • Bacteroidetes
  • Cells, Cultured
  • Desulfovibrio
  • Desulfovibrio vulgaris
  • Dysbiosis / chemically induced
  • Dysbiosis / genetics
  • Dysbiosis / microbiology*
  • Epithelial Cells / drug effects
  • Female
  • Firmicutes
  • Gastrointestinal Microbiome / drug effects*
  • Gene Expression
  • Intestine, Small / cytology
  • Intestine, Small / microbiology
  • Macrophages / drug effects
  • Mice
  • Mice, Inbred C57BL
  • Muramidase / genetics
  • Pancreatitis-Associated Proteins
  • Proteins / genetics
  • RNA, Ribosomal, 16S / analysis*
  • Time Factors
  • Up-Regulation
  • alpha-Defensins / genetics

Substances

  • Anti-Bacterial Agents
  • Antimicrobial Cationic Peptides
  • Atg5 protein, mouse
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Pancreatitis-Associated Proteins
  • Proteins
  • RNA, Ribosomal, 16S
  • Reg3g protein, mouse
  • alpha-Defensins
  • cryptdin 4, mouse
  • Muramidase