Evaluation of Intestinal Dioxin Permeability Using Human Caco-2 Cell Monolayers

Natsume, Yayoi; Satsu, Hideo; Hatsugai, Yasuo; Watanabe, Hisahiko; Sato, Ryuichiro; Ashida, Hitoshi; Tukey, Robert H.; Shimizu, Makoto

doi:10.3136/fstr.9.364

Cited by 8 publications

(9 citation statements)

References 11 publications

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“…In an earlier study the transport across a monolayer of Caco-2 cells of only 2,3,7,8-TCDD has been studied. 56 The authors quantified the TCDD concentration using a luciferase HepG2 cell assay. They found that 15% of TCDD was transported across the Caco-2 cell monolayer in 24 h. 56 This is higher than the 3% transported 2,3,7,8-TCDD observed by us for both models, which might be explained by differences in exposure concentration.…”

Section: Resultsmentioning

confidence: 99%

Implementation of a dynamic intestinal gut-on-a-chip barrier model for transport studies of lipophilic dioxin congeners

et al. 2018

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Section: Resultsmentioning

confidence: 99%

Implementation of a dynamic intestinal gut-on-a-chip barrier model for transport studies of lipophilic dioxin congeners

et al. 2018

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“…Surprisingly, despite the increased CFUs in BHI and Mac plates, the intestinal permeability as measured by FITC-Dextran was not increased (data not shown). In fact, TCDD could decrease the permeability to the Caco-2 cell monolayer in vitro (Natsume et al, 2003). Thus, TCDD could potentially have a beneficial effect on permeability because other AhR ligands increase mucus and anti-microbial peptide production by the host (Thorburn et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

TCDD modulation of gut microbiome correlated with liver and immune toxicity in streptozotocin (STZ)-induced hyperglycemic mice

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Chen

et al. 2016

Toxicology and Applied Pharmacology

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An increasing body of evidence has shown the important role of the gut microbiome in mediating toxicity following environmental contaminant exposure. The goal of this study was to determine if the adverse metabolic effects of chronic 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure would be sufficient to exacerbate hyperglycemia, and to further determine if these outcomes were attributable to the gut microbiota alteration. Adult male CD-1 mice were exposed to TCDD (6 μg/kg body weight biweekly) by gavage and injected (i.p.) with STZ (4 × 50 mg/kg body weight) to induced hyperglycemia. 16S rRNA sequencing was used to characterize the changes in the microbiome community composition. Glucose monitoring, flow cytometry, histopathology, and organ characterization were performed to determine the deleterious phenotypic changes of TCDD exposure. Chronic TCDD treatment did not appear to exacerbate STZ-induced hyperglycemia as blood glucose levels were slightly reduced in the TCDD treated mice; however, polydipsia and polyphagia were observed. Importantly, TCDD exposure caused a dramatic change in microbiota structure, as characterized at the phylum level by increasing Firmicutes and decreasing Bacteroidetes while at the family level most notably by increasing Lactobacillaceae and Desulfovibrionaceae, and decreasing Prevotellaceae and ACK M1. The changes in microbiota were further found to be broadly associated with phenotypic changes seen from chronic TCDD treatment. In particular, the phylum level Bacteroidetes to Firmicutes ratio negatively correlated with both liver weight and liver pathology, and positively associated with %CD3+NK+ T cells, a key mediator of host-microbial interactions. Collectively, these findings suggest that the dysregulated gut microbiome may contribute to the deleterious effects (e.g., liver toxicity) seen with TCDD exposure.

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“…To obtain an experimental system with higher sensitivity and reproducibility, we constructed in the present study a stable dioxinresponsive cell line for a quantitative analysis. Good linearity of the dose-dependent increase in TCDD concentration of the basal solution was obtained in this work (Figure 3), whereas the basal TCDD concentration did not linearly increase when increasing the apical TCDD concentration from zero to 5 nM in the previous study (Natsume et al 2003). The better reproducibility and sensitivity of the present assay system by using stable transfected HepG2 cells enabled us to more accurately evaluate food substances for their TCDDabsorption inhibitory activity.…”

Section: Discussionmentioning

confidence: 58%

“…A differentiated Caco-2 cell monolayer expresses several features of intestinal epithelial cells (Hidalgo et al 1989) and it is widely used as an intestinal epithelial cell model. We have previously attempted to evaluate the amount of TCDD on the basal side of Caco-2 cell monolayers by using HepG2 cells transiently transfected by a plasmid with XRE sequences (Natsume et al 2003). However, this method was not suitable for quantitatively determining the intestinal permeability for TCDD, because the transfection efficiency varied each time.…”

Section: Discussionmentioning

confidence: 99%

“…However, a concise method for assessing dioxin absorption in the small intestine and the effect of food factors on this absorption has not been established. Although we have previously reported an assessment system for this purpose (Natsume et al 2003) by using the AhR-mediated TCDD toxicity expression mechanism just described, a quantitative evaluation of TCDD permeability by this system was not always successful, probably because the transfection efficiency was different among individual wells.…”

Section: Introductionmentioning

confidence: 97%

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In vitro System for Assessing Dioxin Absorption by Intestinal Epithelial Cells and for Preventing this Absorption by Food Substances

et al. 2005

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A system for assessing intestinal dioxin absorption was established by applying a Caco-2 cell monolayer and stable dioxin-responsive cell line. The stable dioxin-responsive cell line was established by introducing a plasmid incorporating the human CYP1A1 promoter into human hepatic HepG2 genomic DNA upstream of the luciferase gene. 2,3,7,8-Tetrachlorodibenzodioxin (TCDD) was added to the apical side of differentiated human intestinal epithelial Caco-2 cell monolayers that had been cultured on a semipermeable membrane. The basal medium was taken after an appropriate incubation time and added to the dioxin-responsive cells, the TCDD content then being analyzed by a luciferase assay. The amount of TCDD in the basal medium increased in a dose- and time-dependent manner, the results being sufficiently sensitive and reproducible. The inhibition of TCDD permeability to the Caco-2 cell monolayer by such food substances as chlorophyll, insoluble corn fiber and tea dregs were observed by this in vitro assessment system. The system will therefore be useful to identify food substances having a preventive effect on the intestinal absorption of dioxins.

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Evaluation of Intestinal Dioxin Permeability Using Human Caco-2 Cell Monolayers

Cited by 8 publications

References 11 publications

Implementation of a dynamic intestinal gut-on-a-chip barrier model for transport studies of lipophilic dioxin congeners

Implementation of a dynamic intestinal gut-on-a-chip barrier model for transport studies of lipophilic dioxin congeners

TCDD modulation of gut microbiome correlated with liver and immune toxicity in streptozotocin (STZ)-induced hyperglycemic mice

In vitro System for Assessing Dioxin Absorption by Intestinal Epithelial Cells and for Preventing this Absorption by Food Substances

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