Intestinal Epithelial Cell Shedding Research Articles

Effects of Schyzocotyle acheilognathi (Yamaguti, 1934) infection on the intestinal microbiota, growth and immune reactions of grass carp (Ctenopharyngodon idella).

Our understanding of interactions among intestinal helminths, gut microbiota and host is still in its infancy in fish. In this study, the effects of Schyzocotyle acheilognathi infection on the intestinal microbiota, growth and immune reactions of grass carp were explored under laboratory conditions. 16S rDNA amplification sequencing results showed that S. acheilognathi infection altered the composition of intestinal microbiota only at the genus level, with a significant increase in the relative abundance of Turicibacter and Ruminococcus (P < 0.05) and a significant decrease in the relative abundance of Gordonia, Mycobacterium and Pseudocanthomonas (P < 0.05). Schyzocotyle acheilognathi infection had no significant effect (P > 0.05) on the alpha diversity indices (including Chao1, ACE, Shannon, Simpson index) of intestinal microbiota in grass carp, but PERMANOVA analysis showed that microbial structure significantly (P < 0.01) differed between hindgut and foregut. PICRUST prediction showed that some metabolism-related pathways were significantly changed after S. acheilognathi infection. The relative abundance of Turicibacter was positively correlated with the fresh weight of tapeworm (foregut: r = 0.48, P = 0.044; hindgut: r = 0.63, P = 0.005). There was no significant difference in the body condition of grass carp between the S. acheilognathi infected group and the uninfected group (P > 0.05). Intestinal tissue section with HE staining showed that S. acheilognathi infection severely damaged the intestinal villi, causing serious degeneration, necrosis and shedding of intestinal epithelial cells. The real-time fluorescent quantitative PCR results showed that S. acheilognathi infection upregulated the mRNA expression of the immune-related genes: Gal1−L2, TGF−β1 and IgM.

Executioner caspases 3 and 7 are dispensable for intestinal epithelium turnover and homeostasis at steady state

Apoptosis is widely believed to be crucial for epithelial cell death and shedding in the intestine, thereby shaping the overall architecture of the gastrointestinal tract, but also regulating tolerance induction, pinpointing a role of apoptosis intestinal epithelial cell (IEC) turnover and maintenance of barrier function, and in maintaining immune homeostasis. To experimentally address this concept, we generated IEC-specific knockout mice that lack both executioner caspase-3 and caspase-7 (Casp3/7ΔIEC), which are the converging point of the extrinsic and intrinsic apoptotic pathway. Surprisingly, the overall architecture, cellular landscape, and proliferation rate remained unchanged in these mice. However, nonapoptotic cell extrusion was increased in Casp3/7ΔIEC mice, compensating apoptosis deficiency, maintaining the same physiological level of IEC shedding. Microbiome richness and composition stayed unaffected, bearing no sign of dysbiosis. Transcriptome and single-cell RNA sequencing analyses of IECs and immune cells revealed no differences in signaling pathways of differentiation and inflammation. These findings demonstrate that during homeostasis, apoptosis per se is dispensable for IEC turnover at the top of intestinal villi intestinal tissue dynamics, microbiome, and immune cell composition.

An IRF1-dependent Pathway of TNFα-induced Shedding in Intestinal Epithelial Cells.

Shedding of intestinal epithelial cells [IECs] is a potent cause of barrier loss which plays an important role in the pathogenesis of inflammatory bowel disease [IBD]. TNFα can induce IEC shedding, but little is known about this process. To investigate the molecular mechanism regulating IEC shedding, mice lacking interferon regulatory factor1 [IRF1], caspase-3, or gasdermin E [GSDME] and their control wild-type [WT] littermates were intravenously injected with tumour necrosis factor alpha [TNFα] to establish an IEC shedding model. A dual-luciferase reporter assay and a chromatin immunoprecipitation assay were used to determine the role of IRF1 in regulating caspase-3 expression. TNFα administration induced obvious IEC shedding in WT mice, but IRF1-/- and caspase-3-/-mice were completely protected from TNFα-induced IEC shedding. As a critical transcription factor, IRF1 was found to be required for caspase-3 expression in IECs by binding to IRF1-binding sites in the caspase-3 promoter. In WT mice, plasma membrane integrity was disrupted in shed IECs; these cells were swollen and contained GSDME-N terminal [NT] fragments which are responsible for the induction of pyroptosis. However, in GSDME-/- mice, plasma membrane integrity was not disrupted in shed IECs, which were not swollen and did not contain GSDME-NT, indicating that GSDME converted TNFα-induced IEC shedding into a pyroptotic cell death process. In addition, IRF1 deficiency resulted in decreases in mucosal inflammation and mucosal bacteria levels in TNFα-challenged colons. IRF1 deficiency maintains intestinal barrier integrity by restricting TNFα-induced IEC shedding.

Intestinal restriction of SalmonellaTyphimurium requires caspase-1 and caspase-11 epithelial intrinsic inflammasomes.

We investigated the role of the inflammasome effector caspases-1 and -11 during Salmonella enterica serovar Typhimurium infection of murine intestinal epithelial cells (IECs). Salmonella burdens were significantly greater in the intestines of caspase-1/11 deficient (Casp1/11−/−), Casp1−/− and Casp11−/− mice, as compared to wildtype mice. To determine if this reflected IEC-intrinsic inflammasomes, enteroid monolayers were derived and infected with Salmonella. Casp11−/− and wildtype monolayers responded similarly, whereas Casp1−/− and Casp1/11−/− monolayers carried significantly increased intracellular burdens, concomitant with marked decreases in IEC shedding and death. Pretreatment with IFN-γ to mimic inflammation increased caspase-11 levels and IEC death, and reduced Salmonella burdens in Casp1−/− monolayers, while high intracellular burdens and limited cell shedding persisted in Casp1/11−/− monolayers. Thus caspase-1 regulates inflammasome responses in IECs at baseline, while proinflammatory activation of IECs reveals a compensatory role for caspase-11. These results demonstrate the importance of IEC-intrinsic canonical and non-canonical inflammasomes in host defense against Salmonella.

Epithelial delamination is protective during pharmaceutical-induced enteropathy

Intestinal epithelial cell (IEC) shedding is a fundamental response to intestinal damage, yet underlying mechanisms and functions have been difficult to define. Here we model chronic intestinal damage in zebrafish larvae using the nonsteroidal antiinflammatory drug (NSAID) Glafenine. Glafenine induced the unfolded protein response (UPR) and inflammatory pathways in IECs, leading to delamination. Glafenine-induced inflammation was augmented by microbial colonization and associated with changes in intestinal and environmental microbiotas. IEC shedding was a UPR-dependent protective response to Glafenine that restricts inflammation and promotes animal survival. Other NSAIDs did not induce IEC delamination; however, Glafenine also displays off-target inhibition of multidrug resistance (MDR) efflux pumps. We found a subset of MDR inhibitors also induced IEC delamination, implicating MDR efflux pumps as cellular targets underlying Glafenine-induced enteropathy. These results implicate IEC delamination as a protective UPR-mediated response to chemical injury, and uncover an essential role for MDR efflux pumps in intestinal homeostasis.

Clostridium perfringens Delta-Toxin Damages the Mouse Small Intestine.

Clostridium perfringens strains B and C cause fatal intestinal diseases in animals. The secreted pore-forming toxin delta-toxin is one of the virulence factors of the strains, but the mechanism of intestinal pathogenesis is unclear. Here, we investigated the effects of delta-toxin on the mouse ileal loop. Delta-toxin caused fluid accumulation and intestinal permeability to fluorescein isothiocyanate (FITC)-dextran in the mouse ileal loop in a dose- and time-dependent manner. Treatment with delta-toxin induced significant histological damage and shortening of villi. Delta-toxin activates a disintegrin and metalloprotease (ADAM) 10, leading to the cleavage of E-cadherin, the epithelial adherens junction protein, in human intestinal epithelial Caco-2 cells. In this study, E-cadherin immunostaining in mouse intestinal epithelial cells was almost undetectable 1 h after toxin treatment. ADAM10 inhibitor (GI254023X) blocked the toxin-induced fluid accumulation and E-cadherin loss in the mouse ileal loop. Delta-toxin stimulated the shedding of intestinal epithelial cells. The shedding cells showed the accumulation of E-cadherin in intracellular vesicles and the increased expression of active caspase-3. Our findings demonstrate that delta-toxin causes intestinal epithelial cell damage through the loss of E-cadherin cleaved by ADAM10.

Bifidobacterium breve reduces apoptotic epithelial cell shedding in an exopolysaccharide and MyD88-dependent manner.

Certain members of the microbiota genus Bifidobacterium are known to positively influence host well-being. Importantly, reduced bifidobacterial levels are associated with inflammatory bowel disease (IBD) patients, who also have impaired epithelial barrier function, including elevated rates of apoptotic extrusion of small intestinal epithelial cells (IECs) from villi—a process termed ‘cell shedding’. Using a mouse model of pathological cell shedding, we show that mice receiving Bifidobacterium breve UCC2003 exhibit significantly reduced rates of small IEC shedding. Bifidobacterial-induced protection appears to be mediated by a specific bifidobacterial surface exopolysaccharide and interactions with host MyD88 resulting in downregulation of intrinsic and extrinsic apoptotic responses to protect epithelial cells under highly inflammatory conditions. Our results reveal an important and previously undescribed role for B. breve, in positively modulating epithelial cell shedding outcomes via bacterial- and host-dependent factors, supporting the notion that manipulation of the microbiota affects intestinal disease outcomes.

Death in the intestinal epithelium-basic biology and implications for inflammatory bowel disease.

Every 4-5 days, intestinal epithelial cells (IEC) are terminated as they reach the end of their life. This process ensures that the epithelium is comprised of the fittest cells that maintain an impermeable barrier to luminal contents and the gut microbiota, as well as the most metabolically able cells that conduct functions in nutrient absorption, digestion, and secretion of antimicrobial peptides. IEC are terminated by apical extrusion-or shedding-from the intestinal epithelial monolayer into the gut lumen. Whether death by apoptosis signals extrusion or death follows expulsion by younger IEC has been a matter of debate. Seemingly a minor detail, IEC death before or after apical extrusion bears weight on the potential contribution of apoptotic IEC to intestinal homeostasis as a consequence of their recognition by intestinal lamina propria phagocytes. In inflammatory bowel disease (IBD), excessive death is observed in the ileal and colonic epithelium. The precise mode of IEC death in IBD is not defined. A highly inflammatory milieu within the intestinal lamina propria, rich in the proinflammatory cytokine, TNF-α, increases IEC shedding and compromises barrier integrity fueling more inflammation. A milestone in the treatment of IBD, anti-TNF-α therapy, may promote mucosal healing by reversing increased and inflammation-associated IEC death. Understanding the biology and consequences of cell death in the intestinal epithelium is critical to the design of new avenues for IBD therapy.

Epidermal growth factor suppresses intestinal epithelial cell shedding through a MAPK-dependent pathway.

Cell shedding from the intestinal villus is a key element of tissue turnover that is essential to maintain health and homeostasis. However, the signals regulating this process are not well understood. We asked whether shedding is controlled by epidermal growth factor receptor (EGFR), an important driver of intestinal growth and differentiation. In 3D ileal enteroid culture and cell culture models (MDCK, IEC-6 and IPEC-J2 cells), extrusion events were suppressed by EGF, as determined by direct counting of released cells or rhodamine-phalloidin labeling of condensed actin rings. Blockade of the MEK-ERK pathway, but not other downstream pathways such as phosphoinositide 3-kinase (PI3K) or protein kinase C (PKC), reversed EGF inhibition of shedding. These effects were not due to a change in cell viability. Furthermore, EGF-driven MAPK signaling inhibited both caspase-independent and -dependent shedding pathways. Similar results were found in vivo, in a novel zebrafish model for intestinal epithelial shedding. Taken together, the data show that EGF suppresses cell shedding in the intestinal epithelium through a selective MAPK-dependent pathway affecting multiple extrusion mechanisms. EGFR signaling might be a therapeutic target for disorders featuring excessive cell turnover, such as inflammatory bowel diseases.

Epigenetic control of intestinal barrier function and inflammation in zebrafish

The intestinal epithelium forms a barrier protecting the organism from microbes and other proinflammatory stimuli. The integrity of this barrier and the proper response to infection requires precise regulation of powerful immune homing signals such as tumor necrosis factor (TNF). Dysregulation of TNF leads to inflammatory bowel diseases (IBD), but the mechanism controlling the expression of this potent cytokine and the events that trigger the onset of chronic inflammation are unknown. Here, we show that loss of function of the epigenetic regulator ubiquitin-like protein containing PHD and RING finger domains 1 (uhrf1) in zebrafish leads to a reduction in tnfa promoter methylation and the induction of tnfa expression in intestinal epithelial cells (IECs). The increase in IEC tnfa levels is microbe-dependent and results in IEC shedding and apoptosis, immune cell recruitment, and barrier dysfunction, consistent with chronic inflammation. Importantly, tnfa knockdown in uhrf1 mutants restores IEC morphology, reduces cell shedding, and improves barrier function. We propose that loss of epigenetic repression and TNF induction in the intestinal epithelium can lead to IBD onset.

Epithelial cell shedding and barrier function: a matter of life and death at the small intestinal villus tip.

The intestinal epithelium is a critical component of the gut barrier. Composed of a single layer of intestinal epithelial cells (IECs) held together by tight junctions, this delicate structure prevents the transfer of harmful microorganisms, antigens, and toxins from the gut lumen into the circulation. The equilibrium between the rate of apoptosis and shedding of senescent epithelial cells at the villus tip, and the generation of new cells in the crypt, is key to maintaining tissue homeostasis. However, in both localized and systemic inflammation, this balance may be disturbed as a result of pathological IEC shedding. Shedding of IECs from the epithelial monolayer may cause transient gaps or microerosions in the epithelial barrier, resulting in increased intestinal permeability. Although pathological IEC shedding has been observed in mouse models of inflammation and human intestinal conditions such as inflammatory bowel disease, understanding of the underlying mechanisms remains limited. This process may also be an important contributor to systemic and intestinal inflammatory diseases and gut barrier dysfunction in domestic animal species. This review aims to summarize current knowledge about intestinal epithelial cell shedding, its significance in gut barrier dysfunction and host-microbial interactions, and where research in this field is directed.

PTU-095 Bifidobacterium Species Reduce Lipopolysaccharide-induced Small Intestinal Epithelial Cell Shedding in Vivo In A Myd88-dependent Manner And Protect Against Dss-induced Colitis

IntroductionCell shedding, the process by which intestinal epithelial cells (IECs) are extruded from the small intestinal (SI) villus is known to be elevated in patients with inflammatory bowel disease (IBD)...

Tu1777 Bifidobacterium Species Reduce Lipopolysaccharide-Induced Small Intestinal Epithelial Cell Shedding In Vivo in a MYD88-Dependent Manner and Protect Against DSS-Induced Colitis

Introduction Cell shedding, the process by which intestinal epithelial cells (IECs) are extruded from the small intestinal (SI) villus is known to be elevated in patients with inflammatory bowel disease (IBD) and is correlated with disease relapse. Importantly, there is evidence that the gut bacterial communities (microbiota) influences intestinal epithelial function including gene expression, cell division and energy balance. We thus sought to determine whether specific members of the microbiota, ‘probiotic’ bifidobacterial species, modulate rates of cell shedding and progression of Dextran sodium sulphate (DSS)-mediated colitis. Methods C57BL/6 mice (WT) or mice deficient in epithelial Myd88 (Vil-Cre +; Myd88 -/-) (Myd88 KO) were orally gavaged with 1 × 10 9 Bifidobacterium breve UCC2003 , B. longum NCIMB8809 or PBS (control) in 3x daily doses. To induce SI cell shedding, mice were injected with 1.25 mg kg-1 Lipopolysaccharide (LPS) intrapertioneally. Animals were euthanized 1.5 hr post-LPS and SI tissue sections analysed for cleaved caspase 3 (CC3) by immunohistochemistry to score shedding along the first 50 cell positions from the villus tip. For colitis studies, control mice or mice colonised with B. breve were administered 2% DSS in drinking water for 6 days and euthanized 8 days post-DSS. Disease activity index (DAI) was recorded daily and histology performed on formalin-fixed tissue sections including periodic acid/Schiff (PAS) stain (goblet cell stain). Results Mice receiving B. breve and B. longum showed less CC3 +ve shedding cells (3.6% +/-0.6, p B. breve was lost in Myd88 KO mice receiving LPS as numbers of CC3 +ve IECs were the same in mice receiving B. breve or vehicle control (13.3%+/-1.7 vs 10.4%+/1.3; ns), indicating that the protective effect may be mediated by Toll-like receptors. In our colitis model, mice colonised with B. breve had reduced DAI compared to control mice, coupled with a significant increase in numbers of PAS +ve goblet cells per crypt (8.2%+/1.6 vs 16.0%+/-0.6; p = 0.05). Conclusion Bifidobacterial species modulate a reduction in rates of cell shedding from the SI villus, potentially via the Myd88 signalling pathway. B. breve is also able to partially ameliorate the adverse effects of DSS-induced colitis through induction of goblet cells. In summary, bifidobacteria, particularly B. breve, may be beneficial as a therapeutic agent for IBD. Disclosure of Interest None Declared.

Inhibition of heat-induced apoptosis in rat small intestine and IEC-6 cells through the AKT signaling pathway

BackgroundAs the world warms up, heat stress is becoming a major cause of economic loss in the livestock industry. Long-time exposure of animals to hyperthermia causes extensive cell apoptosis, which is harmful to them. AKT and AKT-related serine–threonine kinases are known to be involved in signaling cascades that regulate cell survival, but the mechanism remains elusive. In the present study, we demonstrate that phosphoinositide 3-kinase (PI3K) /AKT signal pathway provides protection against apoptosis induced by heat stress to ascertain the key point for treatment.ResultsUnder heat stress, rats showed increased shedding of intestinal epithelial cells. These rats also had elevated levels of serum cortisol and improved expression of heat shock proteins (Hsp27, Hsp70 and Hsp90) in response to heat stress. Apoptosis analysis by TUNEL assay revealed a higher number of villi epithelial cells that were undergoing apoptosis in heat-treated rats than in the normal control. This is supported by gene expression analysis, which showed an increased ratio of Bax/Bcl-2 (p < 0.05), an important indicator of apoptosis. During heat-induced apoptosis, more AKTs were activated, showing increased phosphorylation. An increase of BAD phosphorylation, which is an inhibitory modification, ensued. In rat IEC-6 cell line, a significant higher level of AKT phosphorylation was observed at 2 h after heat exposure. This coincided with a marked reduction of apoptosis.ConclusionTogether, these results suggest that heat stress caused damages to rat jejunum and induced apoptosis to a greater degree. HSPs and pro-survival factors were involved in response to heat stress. Among them, AKT played a key role in inhibiting heat-induced apoptosis.

Substrate Cleavage Profiling Suggests a Distinct Function of Bacteroides fragilis Metalloproteinases (Fragilysin and Metalloproteinase II) at the Microbiome-Inflammation-Cancer Interface

Enterotoxigenic anaerobic Bacteroides fragilis is a significant source of inflammatory diarrheal disease and a risk factor for colorectal cancer. Two distinct metalloproteinase types (the homologous 1, 2, and 3 isoforms of fragilysin (FRA1, FRA2, and FRA3, respectively) and metalloproteinase II (MPII)) are encoded by the B. fragilis pathogenicity island. FRA was demonstrated to be important to pathogenesis, whereas MPII, also a potential virulence protein, remained completely uncharacterized. Here, we, for the first time, extensively characterized MPII in comparison with FRA3, a representative of the FRA isoforms. We employed a series of multiplexed peptide cleavage assays to determine substrate specificity and proteolytic characteristics of MPII and FRA. These results enabled implementation of an efficient assay of MPII activity using a fluorescence-quenched peptide and contributed to structural evidence for the distinct substrate cleavage preferences of MPII and FRA. Our data imply that MPII specificity mimics the dibasic Arg↓Arg cleavage motif of furin-like proprotein convertases, whereas the cleavage motif of FRA (Pro-X-X-Leu-(Arg/Ala/Leu)↓) resembles that of human matrix metalloproteinases. To the best of our knowledge, MPII is the first zinc metalloproteinase with the dibasic cleavage preferences, suggesting a high level of versatility of metalloproteinase proteolysis. Based on these data, we now suggest that the combined (rather than individual) activity of MPII and FRA is required for the overall B. fragilis virulence in vivo.

EGF suppresses intestinal epithelial cell shedding both in vitro and in vivo via a MEK/ERK dependent pathway

Epithelial cell shedding from the intestinal villus is a key element of normal tissue turnover. Increased cell shedding is associated with inflammatory bowel diseases. The signals regulating this process are not well understood. In this study, we asked whether shedding is controlled by epidermal growth factor (EGF) receptor, an important driver of intestinal growth and differentiation.METHODSCell extrusion events in vivo (zebrafish midgut) or in vitro (MDCK or IEC‐6 cells) were identified by rhodamine‐phalloidin labeling of condensed actin rings. In vitro, shedding rates were assessed by counting extruded cells in the culture media over time. EGF, with or without MEK inhibitor (U0126), was given by intraperitoneal injection (fish) or in culture media (cells).RESULTSEGF reduced detectable shedding events on zebrafish villi (34% decrease, p&lt;0.05) and from MDCK (49% decrease, p&lt;0.01) or IEC‐6 (42% decrease, p&lt;0.01) cell monolayers. Pharmacological blockade of MEK/ERK reversed these effects both in vitro and in vivo. In contrast, inhibitors to other EGF‐stimulated pathways such as PI3K/Akt or PKC had no effect.CONCLUSIONSEGF receptor suppresses cell shedding in the intestinal epithelium through a specific MEK/ERK dependent pathway. These results may open the door to new therapeutic avenues for targeting excessive cell turnover in conditions such as inflammatory bowel diseases.

A mouse model of pathological small intestinal epithelial cell apoptosis and shedding induced by systemic administration of lipopolysaccharide

SUMMARYThe gut barrier, composed of a single layer of intestinal epithelial cells (IECs) held together by tight junctions, prevents the entrance of harmful microorganisms, antigens and toxins from the gut lumen into the blood. Small intestinal homeostasis is normally maintained by the rate of shedding of senescent enterocytes from the villus tip exactly matching the rate of generation of new cells in the crypt. However, in various localized and systemic inflammatory conditions, intestinal homeostasis can be disturbed as a result of increased IEC shedding. Such pathological IEC shedding can cause transient gaps to develop in the epithelial barrier and result in increased intestinal permeability. Although pathological IEC shedding has been implicated in the pathogenesis of conditions such as inflammatory bowel disease, our understanding of the underlying mechanisms remains limited. We have therefore developed a murine model to study this phenomenon, because IEC shedding in this species is morphologically analogous to humans. IEC shedding was induced by systemic lipopolysaccharide (LPS) administration in wild-type C57BL/6 mice, and in mice deficient in TNF-receptor 1 (Tnfr1−/−), Tnfr2 (Tnfr2−/−), nuclear factor kappa B1 (Nfκb1−/−) or Nfĸb2 (Nfĸb2−/−). Apoptosis and cell shedding was quantified using immunohistochemistry for active caspase-3, and gut-to-circulation permeability was assessed by measuring plasma fluorescence following fluorescein-isothiocyanate–dextran gavage. LPS, at doses ≥0.125 mg/kg body weight, induced rapid villus IEC apoptosis, with peak cell shedding occurring at 1.5 hours after treatment. This coincided with significant villus shortening, fluid exudation into the gut lumen and diarrhea. A significant increase in gut-to-circulation permeability was observed at 5 hours. TNFR1 was essential for LPS-induced IEC apoptosis and shedding, and the fate of the IECs was also dependent on NFκB, with signaling via NFκB1 favoring cell survival and via NFκB2 favoring apoptosis. This model will enable investigation of the importance and regulation of pathological IEC apoptosis and cell shedding in various diseases.

Shedding-induced gap formation contributes to gut barrier dysfunction in endotoxemia

The intestinal mucosa exhibits high turnover rates with a balance of shedding and the migration of epithelial cells to maintain gut barrier function. Systemic diseases such as sepsis and major thermal injury accelerate the rate of cell shedding, subsequent gap formation, and gut barrier dysfunction. However, the detailed changes of intestinal villi in barrier dysfunction have not been well described. In this study, intestinal barrier dysfunctions were induced through the injection of lipopolysaccharide (LPS) in C57BL/6 mice. Intravital images of the small intestine were observed with multiphoton microscopy for cellular dynamics analysis. The changes of epithelial cells shedding, gap formation, goblet cells, and intestinal leaks were observed, calculated, and analyzed. Endotoxemia enhanced chromatin condensation, accelerated migration, and increased the shedding of intestinal epithelial cells compared with the control group. Furthermore, LPS-induced shedding resulted in gap formation and subsequent intestinal leaks. In total, 40% of intestinal leaks were through gaps, and 60% were through paracellular spaces. Although LPS injection significantly increased the leaks in gaps and paracellular spaces, it did not change the percentage of leaks in gaps and paracellular spaces compared with the control group. We conclude that endotoxemia causes gut barrier dysfunction by increasing epithelium shedding, gaps, and intestinal leaks. However, the effect of the impairment of local barrier maintenance on the distribution of intestinal leaks in gaps and paracellular spaces is minimal.

TNF‐α–induced intestinal epithelial cell shedding: implications for intestinal barrier function

Although epithelial cells are continuously shed from the surface of the intestine, the intestinal epithelium maintains the integrity of the epithelial barrier. This is achieved through a highly dynamic process involving reorganization of tight junction and adherens junction proteins. This process both ejects the cell from the epithelial monolayer and plugs the gap left after the cell is shed. Inflammatory insults can trigger a disturbance of these barrier functions by increasing rates of cell shedding. Epithelial cell shedding and loss of barrier can be visualized by confocal laser endomicroscopy in humans. A simple grading system of confocal laser endomicroscopic images can stratify inflammatory bowel disease (IBD) patients in remission into those who will relapse over the subsequent six months and those who will not. Here, we review the mechanisms governing maintenance of these barrier functions and the implications of inflammation-induced barrier dysfunction in IBD.

PWE-111 Lipopolysaccharide induces small intestinal epithelial cell apoptosis and shedding which is regulated by NFκB signalling

IntroductionThe gut barrier is formed by a single cell thick, simple columnar epithelium consisting of intestinal epithelial cells (IECs) held together by tight junctions. In the small intestine, IECs are...