Chloride channel ClC-2 modulates tight junction barrier function via intracellular trafficking of occludin

2012 ◽  
Vol 302 (1) ◽  
pp. C178-C187 ◽  
Author(s):  
Prashant K. Nighot ◽  
Anthony T. Blikslager
Keyword(s):  
Mass Spectrometry ◽  
Tight Junction ◽  
Chloride Channel ◽  
Barrier Function ◽  
Intracellular Trafficking ◽  
Cell Clone ◽  
Mucosal Barrier ◽  
Caveolin 1 ◽  
Junction Protein ◽  
Mucosal Barrier Function

Previously, we have demonstrated that the chloride channel ClC-2 modulates intestinal mucosal barrier function. In the present study, we investigated the role of ClC-2 in epithelial barrier development and maintenance in Caco-2 cells. During early monolayer formation, silencing of ClC-2 with small interfering (si)RNA led to a significant delay in the development of transepithelial resistance (TER) and disruption of occludin localization. Proteomic analysis employing liquid chromatography-mass spectrometry /mass spectrometry revealed association of ClC-2 with key proteins involved in intracellular trafficking, including caveolin-1 and Rab5. In ClC-2 siRNA-treated cells, occludin colocalization with caveolin-1 was diffuse and in the subapical region. Subapically distributed occludin in ClC-2 siRNA-treated cells showed marked colocalization with Rab5. To study the link between ClC-2 and trafficking of occludin in confluent epithelial monolayers, a Caco-2 cell clone expressing ClC-2 short hairpin (sh)RNA was established. Disruption of caveolae with methyl-β-cyclodextrin (MβCD) caused a marked drop in TER and profound redistribution of caveolin-1-occludin coimmunofluorescence in ClC-2 shRNA cells. In ClC-2 shRNA cells, focal aggregations of Rab5-occludin coimmunofluorescence were present within the cytoplasm. Wortmannin caused an acute fall in TER in ClC-2 shRNA cells and subapical, diffuse redistribution of Rab5-occludin coimmunofluorescence in ClC-2 shRNA cells. An endocytosis and recycling assay for occludin revealed higher basal rate of endocytosis of occludin in ClC-2 shRNA cells. Wortmannin significantly reduced the rate of recycling of occludin in ClC-2 shRNA cells. These data clearly indicate that ClC-2 plays an important role in the modulation of tight junctions by influencing caveolar trafficking of the tight junction protein occludin.

scholarly journals Konjac glucomannan polysaccharide and inulin oligosaccharide enhance the colonic mucosal barrier function and modulate gut-associated lymphoid tissue immunity in C57BL/6J mice

2019 ◽  
Vol 123 (3) ◽  
pp. 319-327 ◽  
Author(s):  
Chih-Hsuan Changchien ◽  
Yi-Chun Han ◽  
Hsiao-Ling Chen
Keyword(s):  
Gene Expression ◽  
Tight Junction ◽  
Barrier Function ◽  
Control Diet ◽  
Konjac Glucomannan ◽  
Mucosal Barrier ◽  
Low Level ◽  
Inflammatory Status ◽  
Junction Protein ◽  
Mucosal Barrier Function

AbstractBoth konjac glucomannan (KGM) and inulin oligosaccharide have been shown to improve bowel function, but their effects on the mucosal barrier function and immunity are not fully understood. The aim of the present study was to determine the effects of a low-level supplementation of dietary fibres on the colonic mucosal barrier function, antioxidant enzyme defence and immunity. C57BL/6J mice (6 weeks of age, eight per group) were randomly assigned to consume one of the following diets: control or control diet supplemented with 2 % (w/w) of KGM, inulin oligosaccharide (degree polymerisation = 8) or KGM+inulin (1 %, w/w each (K+I)). Fresh faeces were collected on days 19–21. Mice were killed on day 22 after fasting. Segments of colon tissues were processed for histological procedure and stained for acidic mucins and tight junction protein marker zona occludin-1 (ZO-1). The remaining tissues were processed to determine the gene expression of mucin 2, tight junction proteins, antioxidant enzymes and cytokines. The plasma cytokines were measured. Results indicated that KGM, inulin and K+I significantly increased the mucosal layer thickness, mucin density (granule number/crypt) and gene expression of Muc2 as compared with the control. All fibre treatments increased the gene expressions of ZO-1, occludin, glutathione peroxidase, glutathione S-transferase π, catalase and IL-10. In addition, all fibre treatments increased faecal butyrate and probiotics, and plasma IL-10 concentrations. In conclusion, supplementation of low-level, 2 % (w/w), of K+I was sufficient to enhance the mucosal barrier function and anti-inflammatory status.

scholarly journals Th17 Cytokines Disrupt the Airway Mucosal Barrier in Chronic Rhinosinusitis

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mahnaz Ramezanpour ◽  
Sophia Moraitis ◽  
Jason L. P. Smith ◽  
P. J. Wormald ◽  
Sarah Vreugde
Keyword(s):  
Chronic Rhinosinusitis ◽  
Barrier Function ◽  
Paracellular Permeability ◽  
Mucosal Barrier ◽  
Human Nasal Epithelial Cells ◽  
Basolateral Side ◽  
Junction Protein ◽  
Mucosal Barrier Function ◽  
Th17 Cytokines ◽  
Zona Occludens

Cytokine mediated changes in paracellular permeability contribute to a multitude of pathological conditions including chronic rhinosinusitis (CRS). The purpose of this study was to investigate the effect of interferons and of Th1, Th2, and Th17 cytokines on respiratory epithelium barrier function. Cytokines and interferons were applied to the basolateral side of air-liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) from CRS with nasal polyp patients. Transepithelial electrical resistance (TEER) and permeability of FITC-conjugated dextrans were measured over time. Additionally, the expression of the tight junction protein Zona Occludens-1 (ZO-1) was examined via immunofluorescence. Data was analysed using ANOVA, followed by Tukey HSD post hoc test. Our results showed that application of interferons and of Th1 or Th2 cytokines did not affect the mucosal barrier function. In contrast, the Th17 cytokines IL-17, IL-22, and IL-26 showed a significant disruption of the epithelial barrier, evidenced by a loss of TEER, increased paracellular permeability of FITC-dextrans, and discontinuous ZO-1 immunolocalisation. These results indicate that Th17 cytokines may contribute to the development of CRSwNP by promoting a leaky mucosal barrier.

scholarly journals Dietary fibre affects intestinal mucosal barrier function and regulates intestinal bacteria in weaning piglets

2013 ◽  
Vol 110 (10) ◽  
pp. 1837-1848 ◽  
Author(s):  
Hong Chen ◽  
Xiangbing Mao ◽  
Jun He ◽  
Bing Yu ◽  
Zhiqing Huang ◽  
...  
Keyword(s):  
Barrier Function ◽  
Transforming Growth Factor ◽  
Control Diet ◽  
Mucosal Barrier ◽  
Intestinal Barrier Function ◽  
Mrna Levels ◽  
Intestinal Mucosal Barrier ◽  
Junction Protein ◽  
Mucosal Barrier Function ◽  
Weaning Piglets

The objective of the present study was to evaluate the effects of fibre source on intestinal mucosal barrier function in weaning piglets. A total of 125 piglets were randomly allotted on the basis of their body weight and litters to one of five experimental diets, i.e. a control diet without fibre source (CT), and diets in which expanded maize was replaced by 10 % maize fibre (MF), 10 % soyabean fibre (SF), 10 % wheat bran fibre (WBF) or 10 % pea fibre (PF). The diets and water were fed ad libitum for 30 d. Piglets on the WBF and PF diets had lower diarrhoea incidence compared with the MF- and SF-fed animals. A higher ratio of villous height:crypt depth in the ileum of WBF-fed piglets and higher colonic goblet cells in WBF- and PF-fed piglets were observed compared with CT-, MF- and SF-fed piglets. In the intestinal digesta, feeding WBF and PF resulted in increased Lactobacillus counts in the ileum and Bifidobacterium counts in the colon. Lower Escherichia coli counts occurred in the ileum and colon of WBF-fed piglets than in SF-fed piglets. Tight junction protein (zonula occludens 1; ZO-1) and Toll-like receptor 2 (TLR2) gene mRNA levels were up-regulated in the ileum and colon of pigs fed WBF; however, feeding MF and SF raised IL-1α and TNF-α mRNA levels. Furthermore, higher diamine oxidase activities, transforming growth factor-α, trefoil factor family and MHC-II concentration occurred when feeding WBF and PF. In conclusion, the various fibre sources had different effects on the ileal and colonic barrier function. Clearly, WBF and PF improved the intestinal barrier function, probably mediated by changes in microbiota composition and concomitant changes in TLR2 gene expression.

scholarly journals A Role of Exopolysaccharide Produced by Streptococcus thermophilus in the Intestinal Inflammation and Mucosal Barrier in Caco-2 Monolayer and Dextran Sulphate Sodium-Induced Experimental Murine Colitis

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 513 ◽  
Author(s):  
Yun Chen ◽  
Ming Zhang ◽  
Fazheng Ren
Keyword(s):  
Tight Junction ◽  
Intestinal Inflammation ◽  
Gastrointestinal Disease ◽  
Streptococcus Thermophilus ◽  
Water Soluble ◽  
Mucosal Barrier ◽  
Dextran Sulphate ◽  
Dextran Sulphate Sodium ◽  
Junction Protein ◽  
Mucosal Barrier Function

Exopolysaccharide (EPS) produced by probiotics may play an important role in gastrointestinal disease prevention, including ulcerative colitis. However, there is no literature reporting on the intervention effects of purified EPS. The aim of this study was to investigate the alleviating effect of the purified EPS produced by Streptococcus thermophilus MN-BM-A01 on murine model of colitis induced by dextran sulphate sodium (DSS). A water-soluble heteropolysaccharide (EPS-1) isolated from MN-BM-A01 was composed of rhamnose, glucose, galactose, and mannose in a molar ratio of 12.9:26.0:60.9:0.25, with molecular weight of 4.23 × 105 Da. After EPS-1 administration, the disease severity of mouse colitis was significantly alleviated, mainly manifesting as the decrease of disease activity index and mitigated colonic epithelial cell injury. Meanwhile, pro-inflammatory cytokines levels (tumor necrosis factor-α, interleukin-6, and interferon-γ) were significantly suppressed, the reduced expressions of tight junction protein (claudin-1, occludin, and E-canherin) were counteracted. In addition, the results in vitro showed that EPS-1 protected intestinal barrier integrity from the disruption by lipopolysaccharide in Caco-2 monolayer, increased expression of tight junction and alleviated pro-inflammatory response. Collectively, our study confirmed the protective effects of purified EPS produced by Streptococcus thermophilus on acute colitis via alleviating intestinal inflammation and improving mucosal barrier function.

Methamphetamine-induced alterations in intestinal mucosal barrier function occur via the microRNA-181c/ TNF-α/tight junction axis

2020 ◽  
Vol 321 ◽  
pp. 73-82 ◽  
Author(s):  
Simin Shen ◽  
Jingjiao Zhao ◽  
Yicong Dai ◽  
Fengrong Chen ◽  
Zunyue Zhang ◽  
...  
Keyword(s):  
Tight Junction ◽  
Barrier Function ◽  
Mucosal Barrier ◽  
Intestinal Mucosal Barrier ◽  
Tnf Α ◽  
Mucosal Barrier Function

scholarly journals Painong San, a Traditional Chinese Compound Herbal Medicine, Restores Colon Barrier Function on DSS-Induced Colitis in Mice

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Xuelin Rui ◽  
Jiacheng Li ◽  
Ye Yang ◽  
Li Xu ◽  
Yang Liu ◽  
...  
Keyword(s):  
Herbal Medicine ◽  
Tight Junction ◽  
Barrier Function ◽  
Notch Pathway ◽  
The Body ◽  
Mucosal Barrier ◽  
Control Group ◽  
Mucosal Permeability ◽  
Mucosal Barrier Function ◽  
Related Proteins

Objective. The intestinal barrier decreases in colitis and restores the integrity of the mucosal barriers that could be used for the treatment of colitis. Painong San (PNS), a traditional Chinese compound herbal medicine originally recorded in “Jingui Yaolve” by Zhongjing Zhang in the Later Han Dynasty, is often used in China and Japan to treat various purulent diseases including intestinal carbuncle. This study was to investigate the effect of PNS on mucosal barrier function in mice with DSS-induced colitis and its related mechanisms. Methods. BALB/C mice were given 3% DSS to induce colitis. The body weight and stool status of the mice were recorded daily, and the histopathological changes of the colon were observed after execution. The permeability of the intestinal mucosa was measured by fluorescein isothiocyanate-dextran 4000, the change of intestinal microbiota was measured by 16S rDNA, and the tight junction-related proteins and Muc-2 were investigated by immunohistochemical or immunofluorescence. The possible signaling pathways were detected by western blot. Results. Compared with the control group, the composition of the microbiota in the PNS group was close to that of the normal group, the number of goblet cells was improved, and the mucosal permeability was significantly reduced. PNS could upregulate the expression of tight junction-related proteins (ZO-1, claudin-1, and occludin) and Muc-2, and at the same time, regulate the Notch pathway. Conclusion. PNS could effectively improve the mucosal barrier function through multiple ways, including restoring the balance of intestine flora, enhancement of the mucous layer barrier, and mechanical barrier function. These protective effects may relate to inhibiting the Notch signaling pathway activated by DSS.

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