Intestinal Epithelial Monolayer Permeability of Sweet Potato-Derived Anthocyanin and Carotenoid Extracts in Caco-2 Cells

Background: Dysfunction of the intestinal mucosal barrier plays an important role in the pathophysiology of severe acute pancreatitis (SAP). Continuous blood purification (CBP) has been shown to improve the prognosis of SAP patients. In order to investigate the effect of CBP on intestinal mucosal barrier dysfunction in SAP patients with MODS, we conducted in vivo and in vitro experiments to explore the underlying mechanisms. Methods: The markers for the assessment of intestinal mucosal barrier function including serum diamine oxidase (DAO), endotoxin and intestinal epithelial monolayer permeability were detected during CBP therapy. The distribution and expression of cytoskeleton protein F-actin and tight junction proteins claudin-1 were observed. In addition, Rho kinase (ROCK) mRNA expression and serum tumor necrosis factor-alpha (TNF-α) levels during CBP were determined. Results: SAP patients with MODS had increased levels of serum DAO, endotoxin and intestinal epithelial monolayer permeability when compared with normal controls. While the distribution of F-actin and claudin-1 was rearranged, and the expression of claudin-1 significantly decreased, but F-actin had no change. Meanwhile, ROCK mRNA expression and serum TNF-α level were increased. However, after CBP treatment, levels of serum DAO, endotoxin and intestinal epithelial monolayer permeability decreased. The F-actin and claudin-1 reorganization attenuated and the expression of claudin-1 increased. At the same time, ROCK mRNA expression and serum TNF-α level were decreased. Conclusions: CBP can effectively improve intestinal mucosal barrier dysfunction. The beneficial effect is associated with the improvement of cytoskeleton and tight junction proteins in stability by downregulation of ROCK mRNA expression through the removal of excess proinflammatory factors.

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Inhibition of Matriptase Activity Results in Decreased Intestinal Epithelial Monolayer Integrity In Vitro

PLoS ONE ◽

10.1371/journal.pone.0141077 ◽

2015 ◽

Vol 10 (10) ◽

pp. e0141077 ◽

Cited By ~ 4

Author(s):

E. Pászti-Gere ◽

S. McManus ◽

N. Meggyesházi ◽

P. Balla ◽

P. Gálfi ◽

...

Keyword(s):

Intestinal Epithelial ◽

Epithelial Monolayer

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Octreotide Alleviates Autophagy by Up-Regulation of MicroRNA-101 in Intestinal Epithelial Cell Line Caco-2

Cellular Physiology and Biochemistry ◽

10.1159/000493413 ◽

2018 ◽

Vol 49 (4) ◽

pp. 1352-1363 ◽

Cited By ~ 5

Author(s):

Yuling Li ◽

Su Wang ◽

Xingjuan Gao ◽

Ying Zhao ◽

Yongwei Li ◽

...

Keyword(s):

Tight Junction ◽

Cell Viability ◽

Mtor Pathway ◽

Common Side Effect ◽

Epithelial Cell Line ◽

Therapeutic Effects ◽

Intestinal Epithelial ◽

Differentiated Cells ◽

Monolayer Permeability ◽

Underlying Mechanisms

Background: Intestinal mucositis is a common side-effect after anti-cancer therapy, which may greatly restrict the therapeutic effects. We aimed to explore the functional role of octreotide (OCT) in lipopolysaccharide (LPS)-induced autophagy of human intestinal epithelial cells as well as the underlying mechanisms. Methods: Cell viability and expression of proteins related to autophagy, AMPK and the mTOR pathway in LPS-treated Caco-2 cells were determined by CCK-8 assay and Western blot analysis, respectively. Effects of OCT on LPS-induced alterations as well as miR-101 expression were measured. Then, miR-101 was aberrantly expressed, and whether OCT alleviated LPS-induced autophagy through miR-101 was tested. Next, whether TGF-β-activated kinase 1 (TAK1) was involved in the regulation of miR-101 in LPS-induced autophagy was studied. Effects of OCT on monolayer permeability and tight junction level were analyzed via measuring transepithelial electrical resistance (TEER) and expression of tight junction proteins. Results: LPS reduced cell viability and increased autophagy through activating AMPK and inhibiting the mTOR pathway in Caco-2 cells. OCT alleviated LPS-induced alterations and repressed degradation of autophagosome. Then, we found that OCT affected autophagy through up-regulating miR-101 in LPS-treated cells. Moreover, miR-101-induced inactivation of AMPK and activation of the mTOR pathway in LPS-treated cells were reversed by inhibition of TAK1 phosphorylation. Finally, we found miR-101 was up-regulated in differentiated cells, and OCT protected the monolayer permeability and tight junction level. Conclusion: OCT repressed autophagy through miR-101-mediated inactivation of TAK1, along with inactivation of AMPK and activation of the mTOR pathway in LPS-treated Caco-2 cells.

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Reversible disassembly of an intestinal epithelial monolayer by prolonged exposure to phorbol ester

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1994.266.2.g214 ◽

1994 ◽

Vol 266 (2) ◽

pp. G214-G221 ◽

Cited By ~ 8

Author(s):

G. Hecht ◽

B. Robinson ◽

A. Koutsouris

Keyword(s):

Tight Junction ◽

Phorbol Ester ◽

Barrier Function ◽

Prolonged Exposure ◽

Morphological Changes ◽

Cell Types ◽

Intestinal Epithelial ◽

Epithelial Monolayer ◽

Epithelial Barrier Function ◽

Functional Changes

This article describes a model of reversible disassembly of a cultured human intestinal epithelial monolayer by prolonged exposure to the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). Prolonged phorbol ester exposure reduces protein kinase C (PKC) levels in numerous cell types including T84, as shown here. Under PKC-downregulated conditions, T84 monolayers, which simulate the highly organized structure of native intestinal crypt cells, become disassembled into 2 or 3 layers of rounded cells. Proliferation does not account for these morphological changes as assessed by thymidine incorporation studies. The effects of structural disorganization on epithelial barrier function was also examined. The permeability of disassembled monolayers was significantly greater than that of controls. Flux studies localized the permeability defect to the tight junction. PKC-associated alterations in the perijunctional ring of actin and myosin, one of the putative regulators of flow across the tight junction, were found to correlate with the observed functional changes. Most interesting was the fact that monolayer reassembly to the original columnar epithelial phenotype and reestablishment of barrier function occurred upon normalization of PKC levels. This model of reversible monolayer disassembly will allow investigation into the relationship between epithelial structure and function and examination of factors that govern monolayer formation.

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Active and Repressive Chromatin-Associated Proteome after MPA Treatment and the Role of Midkine in Epithelial Monolayer Permeability

International Journal of Molecular Sciences ◽

10.3390/ijms17040597 ◽

2016 ◽

Vol 17 (4) ◽

pp. 597 ◽

Cited By ~ 3

Author(s):

Niamat Khan ◽

Christof Lenz ◽

Lutz Binder ◽

Dasaradha Pantakani ◽

Abdul Asif

Keyword(s):

Epithelial Monolayer ◽

Repressive Chromatin ◽

Monolayer Permeability

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Effect of Ciprofloxacin on Shiga Toxin Production and Translocation across an Intestinal Epithelial Monolayer

Pediatric Research ◽

10.1203/00006450-199904020-00971 ◽

1999 ◽

Vol 45 (4, Part 2 of 2) ◽

pp. 163A-163A

Author(s):

Jenifer L Jaeger ◽

David Acheson

Keyword(s):

Shiga Toxin ◽

Toxin Production ◽

Intestinal Epithelial ◽

Epithelial Monolayer

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Murine Norovirus Transcytosis across anIn VitroPolarized Murine Intestinal Epithelial Monolayer Is Mediated by M-Like Cells

Journal of Virology ◽

10.1128/jvi.02378-13 ◽

2013 ◽

Vol 87 (23) ◽

pp. 12685-12693 ◽

Cited By ~ 31

Author(s):

Mariam B. Gonzalez-Hernandez ◽

Thomas Liu ◽

Luz P. Blanco ◽

Heather Auble ◽

Hilary C. Payne ◽

...

Keyword(s):

Viral Entry ◽

Cell Monolayer ◽

Small Animal ◽

Intestinal Epithelial ◽

Murine Norovirus ◽

Small Animal Model ◽

Intestinal Epithelial Barrier ◽

M Cell ◽

Epithelial Monolayer

Noroviruses (NoVs) are the causative agent of the vast majority of nonbacterial gastroenteritis worldwide. Due to the inability to culture human NoVs and the inability to orally infect a small animal model, little is known about the initial steps of viral entry. One particular step that is not understood is how NoVs breach the intestinal epithelial barrier. Murine NoV (MNV) is the only NoV that can be propagatedin vitroby infecting murine macrophages and dendritic cells, making this virus an attractive model for studies of different aspects of NoV biology. Polarized murine intestinal epithelial mICcl2cells were used to investigate how MNV interacts with and crosses the intestinal epithelium. In thisin vitromodel of the follicle-associated epithelium (FAE), MNV is transported across the polarized cell monolayer in the absence of viral replication or disruption of tight junctions by a distinct epithelial cell with microfold (M) cell properties. In addition to transporting MNV, these M-like cells also transcytose microbeads and express an IgA receptor. Interestingly, B myeloma cells cultured in the basolateral compartment underlying the epithelial monolayer did not alter the number of M-like cells but increased their transcytotic activity. Our data demonstrate that MNV can cross an intact intestinal epithelial monolayerin vitroby hijacking the M-like cells' intrinsic transcytotic pathway and suggest a potential mechanism for MNV entry into the host.

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