Obesity-Linked Gut Microbiome Dysbiosis Associated with Derangements in Gut Permeability and Intestinal Cellular Homeostasis Independent of Diet

Ravinder Nagpal, Tiffany M Newman, Shaohua Wang, Shalini Jain, James F Lovato, Hariom Yadav, Ravinder Nagpal, Tiffany M Newman, Shaohua Wang, Shalini Jain, James F Lovato, Hariom Yadav

Abstract

This study aimed to determine the association between non-high-fat diet-induced obesity- (non-DIO-) associated gut microbiome dysbiosis with gut abnormalities like cellular turnover of intestinal cells, tight junctions, and mucin formation that can impact gut permeability. We used leptin-deficient (Lepob/ob) mice in comparison to C57BL/6J control mice, which are fed on identical diets, and performed comparative and correlative analyses of gut microbiome composition, gut permeability, intestinal structural changes, tight junction-mucin formation, cellular turnover, and stemness genes. We found that obesity impacted cellular turnover of the intestine with increased cell death and cell survival/proliferation gene expression with enhanced stemness, which are associated with increased intestinal permeability, changes in villi/crypt length, and decreased expression of tight junctions and mucus synthesis genes along with dysbiotic gut microbiome signature. Obesity-induced gut microbiome dysbiosis is also associated with abnormal intestinal organoid formation characterized with decreased budding and higher stemness. Results suggest that non-DIO-associated gut microbiome dysbiosis is associated with changes in the intestinal cell death versus cell proliferation homeostasis and functions to control tight junctions and mucous synthesis-regulating gut permeability.

Figures

Figure 1
Figure 1
Gut permeability and intestinal structural changes are associated with mucin-tight junction formation independent of dietary differences. (a) Gut permeability (diffusion of FITC-dextran; 4 KDa from gut to blood) increased obese mice compared to B6 control. (b) Expression of mucin (Muc2 and Muc6) and tight junction- (occludin-, Zo1-, and Jam-) forming genes significantly decreased in Lepob/ob intestine than B6 mice. (c, d) Western blot analyses of Zo1, occludin, and tubulin in intestinal tissues. (e–g) Hematoxylin and eosin (20x) staining for the small intestine (ileum) (e), decreased length of villi (f), and increased crypt (g) in obese mice. Values presented here are average ± SEM. p values are defined as ∗ < 0.05, ∗∗ < 0.01, and ∗∗∗ < 0.001.
Figure 2
Figure 2
Non-diet-induced obesity developed stable changes in cells to form abnormal intestinal structures and influenced intestinal cellular homeostasis. (a–c) Intestinal organoids (a) and their budding potential (b, c) decreased in Lepob/ob-derived organoids versus B6 mice. (d–i) mRNA expression of cell death (Bad, Bax, and Bcl2), cell proliferation (Ccnd1, Cdk6, and Sox4), and stem cell- (Lgr5-, Olfm4-, and Bmi1-) specific genes in obese and B6 intestines. Values presented here are average ± SEM/SD. p values are defined as ∗ < 0.05, ∗∗ < 0.01, and ∗∗∗ < 0.001.
Figure 3
Figure 3
Lepob/ob mice exhibit significant gut microbiome dysbiosis compared to C57BL/6J (B6) mice fed with identical diet. (a) PCoA analysis shows differential clustering of the microbiome signature in Lepob/ob and B6 mice. (b, c) Microbial phylum abundance and Firmicutes : Bacteroidetes (F : B) ratio in Lepob/ob and B6 mice. (d–f) Significantly differing microbial families (d) and genera (e) abundances as well as fold change (percent) between Lepob/ob and B6 mice. Values presented here are average ± SEM/SD.
Figure 4
Figure 4
Relationship of metabolic functions, intestinal morphology, and functions with gut microbiome dysbiosis, independent of dietary differences. Spearman correlation analysis to establish the association between metabolic measures (body weight, blood glucose, AUC-GTT, and AUC-ITT), gut permeability changes, intestinal structural changes, and gene expression changes and the gut microbiome signature between Lepob/ob and B6 mice. Values presented here are average of 3–8 replicates. p values are defined as ∗ < 0.05 and ∗∗ < 0.01.

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Source: PubMed

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