Effect of curcumin on acidic pH-induced expression of IL-6 and IL-8 in human esophageal epithelial cells (HET-1A): role of PKC, MAPKs, and NF-κB

2009 ◽  
Vol 296 (2) ◽  
pp. G388-G398 ◽  
Author(s):  
Parvaneh Rafiee ◽  
Victoria M. Nelson ◽  
Sharon Manley ◽  
Michael Wellner ◽  
Martin Floer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mucosal Injury ◽  
Inhibitory Effect ◽  
Acidic Ph ◽  
Induced Expression ◽  
Potent Inducer ◽  
Mrna And Protein Expression ◽  
Esophageal Epithelial Cells ◽  
P38 Mapk Inhibitor

Human esophageal epithelial cells play a key role in esophageal inflammation in response to acidic pH during gastroesophageal reflux disease (GERD), increasing secretion of IL-6 and IL-8. The mechanisms underlying IL-6 and IL-8 expression and secretion in esophageal epithelial cells after acid stimulation are not well characterized. We investigated the role of PKC, MAPK, and NF-κB signaling pathways and transcriptional regulation of IL-6 and IL-8 expression in HET-1A cells exposed to acid. Exposure of HET-1A cells to pH 4.5 induced NF-κB activity and enhanced IL-6 and IL-8 secretion and mRNA and protein expression. Acid stimulation of HET-1A cells also resulted in activation of MAPKs and PKC (α and ε). Curcumin, as well as inhibitors of NF-κB (SN-50), PKC (chelerythrine), and p44/42 MAPK (PD-098059) abolished the acid-induced expression of IL-6 and IL-8. The JNK inhibitor SP-600125 blocked expression/secretion of IL-6 but only partially attenuated IL-8 expression. The p38 MAPK inhibitor SB-203580 did not inhibit IL-6 expression but exerted a stronger inhibitory effect on IL-8 expression. Together, these data demonstrate that 1) acid is a potent inducer of IL-6 and IL-8 production in HET-1A cells; 2) MAPK and PKC signaling play a key regulatory role in acid-mediated IL-6 and IL-8 expression via NF-κB activation; and 3) the anti-inflammatory plant compound curcumin inhibits esophageal activation in response to acid. Thus IL-6 and IL-8 expression by acid may contribute to the pathobiology of mucosal injury in GERD, and inhibition of the NF-κB/proinflammatory cytokine pathways may emerge as important therapeutic targets for treatment of esophageal inflammation.

Su1103 The Role of Serotonin in Reflux-Induced Esophageal Mucosal Injury in Human Esophageal Epithelial Cells

2016 ◽  
Vol 150 (4) ◽  
pp. S470
Author(s):  
Liping Wu ◽  
Tadayuki Oshima ◽  
Yoshio Ohda ◽  
Toshihiko Tomita ◽  
Hirokazu Fukui ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mucosal Injury ◽  
Esophageal Epithelial Cells

Effect of Calcium on the Growth and Differentiation of Cultured Rat Esophageal Epithelial Cells

1982 ◽  
Vol 40 ◽  
pp. 132-133
Author(s):  
W.T. Gunning ◽  
M.R. Marino ◽  
M.S. Babcock ◽  
G.D. Stoner
Keyword(s):  
Epithelial Cells ◽  
Cell Types ◽  
Replication Rate ◽  
Enzymatic Dissociation ◽  
Growth Assay ◽  
Epidermal Growth ◽  
Fetal Bovine ◽  
Esophageal Epithelial Cells ◽  
Cellular Replication

The role of calcium in modulating cellular replication and differentiation has been described for various cell types. In the present study, the effects of Ca++ on the growth and differentiation of cultured rat esophageal epithelial cells was investigated.Epithelial cells were isolated from esophagi taken from 8 week-old male CDF rats by the enzymatic dissociation method of Kaighn. The cells were cultured in PFMR-4 medium supplemented with 0.25 mg/ml dialyzed fetal bovine serum, 5 ng/ml epidermal growth factor, 10-6 M hydrocortisone 10-6 M phosphoethanolamine, 10-6 M ethanolamine, 5 pg/ml insulin, 5 ng/ml transferrin, 10 ng/ml cholera toxin and 50 ng/ml garamycin at 36.5°C in a humidified atmosphere of 3% CO2 in air. At weekly intervals, the cells were subcultured with a solution containing 1% polyvinylpyrrolidone, 0.01% EGTA, and 0.05% trypsin. After various passages, the replication rate of the cells in PFMR-4 medium containing from 10-6 M to 10-3 M Ca++ was determined using a clonal growth assay.

scholarly journals Functional role of kallikrein 5 and proteinase-activated receptor 2 in eosinophilic esophagitis

2020 ◽  
Vol 12 (545) ◽  
pp. eaaz7773 ◽  
Author(s):  
Nurit P. Azouz ◽  
Andrea M. Klingler ◽  
Purnima Pathre ◽  
John A. Besse ◽  
Netali Ben Baruch-Morgenstern ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Eosinophilic Esophagitis ◽  
Cytokine Production ◽  
Clinical Samples ◽  
Microbiome Composition ◽  
Peptidase Inhibitor ◽  
Esophageal Epithelial Cells ◽  
Protease Activated Receptor 2

Eosinophilic esophagitis (EoE) is a chronic, food antigen–driven, inflammatory disease of the esophagus and is associated with impaired barrier function. Evidence is emerging that loss of esophageal expression of the serine peptidase inhibitor, kazal type 7 (SPINK7), is an upstream event in EoE pathogenesis. Here, we provide evidence that loss of SPINK7 mediates its pro-EoE effects via kallikrein 5 (KLK5) and its substrate, protease-activated receptor 2 (PAR2). Overexpression of KLK5 in differentiated esophageal epithelial cells recapitulated the effect of SPINK7 gene silencing, including barrier impairment and loss of desmoglein-1 expression. Conversely, KLK5 deficiency attenuated allergen-induced esophageal protease activity, modified commensal microbiome composition, and attenuated eosinophilia in a murine model of EoE. Inhibition of PAR2 blunted the cytokine production associated with loss of SPINK7 in epithelial cells and attenuated the allergen-induced esophageal eosinophilia in vivo. Clinical samples substantiated dysregulated PAR2 expression in the esophagus of patients with EoE, and delivery of the clinically approved drug α1 antitrypsin (A1AT, a protease inhibitor) inhibited experimental EoE. These findings demonstrate a role for the balance between KLK5 and protease inhibitors in the esophagus and highlight EoE as a protease-mediated disease. We suggest that antagonizing KLK5 and/or PAR2 has potential to be therapeutic for EoE.

scholarly journals Metformin Stimulates FGF21 Expression in Primary Hepatocytes

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Eva B. Nygaard ◽  
Sara G. Vienberg ◽  
Cathrine Ørskov ◽  
Harald S. Hansen ◽  
Birgitte Andersen
Keyword(s):  
Human Hepatocytes ◽  
Fibroblast Growth Factor 21 ◽  
Potent Inducer ◽  
Glucose And Lipid Metabolism ◽  
Compound C ◽  
Mrna And Protein Expression ◽  
Dose Dependent Increase ◽  
Fgf21 Expression

Fibroblast growth factor 21 (FGF21) is a novel metabolic regulator of glucose and lipid metabolism; however, the exact mechanism of action and regulation of FGF21 is not fully understood. Metabolic status plays an important role in the regulation of FGF21, and we therefore examined whether metformin, an indirect AMPK-activator, regulates FGF21 expression in hepatocytes. FGF21 mRNA and protein expression were determined after incubation of primary cultured rat and human hepatocytes with metformin for 24 hours. To study the role of AMPK in the putative regulation of FGF21, hepatocytes were incubated with Compound C (an AMPK inhibitor) in the presence of metformin. A strong dose-dependent increase in FGF21 expression was observed in both rat and human hepatocytes treated with metformin. This effect was blocked by addition of the AMPK-inhibitor Compound C. The study shows that metformin is a potent inducer of hepatic FGF21 expression and that the effect of metformin seems to be mediated through AMPK activation. As FGF21 therapy normalizes blood glucose in animal models of type 2 diabetes, the induction of hepatic FGF21 by metformin might play an important role in metformin’s antidiabetic effect.

scholarly journals IL-27 Modulates Chemokine Production in TNF-α -Stimulated Human Oral Epithelial Cells

2017 ◽  
Vol 43 (3) ◽  
pp. 1198-1206 ◽  
Author(s):  
Yoshitaka Hosokawa ◽  
Ikuko Hosokawa ◽  
Kazumi Ozaki ◽  
Takashi Matsuo
Keyword(s):  
Signal Transduction ◽  
Epithelial Cells ◽  
Inhibitory Effect ◽  
Chemokine Production ◽  
Oral Epithelial Cells ◽  
Phosphorylation Level ◽  
Stat3 Phosphorylation ◽  
Tnf Α ◽  
Oral Epithelial

Background/Aims: Interleukin-27 (IL-27) is a cytokine which belongs to the IL-12 family. However, the role of IL-27 in the pathogenesis of periodontal disease is uncertain. The aim of this study was to examine the effect of IL-27 on chemokine production in TNF-α-stimulated human oral epithelial cells (TR146). Methods: We measured chemokine production in TR146 by ELISA. We used western blot analysis to detect the phosphorylation levels of signal transduction molecules, including STAT1 and STAT3 in TR146. We used inhibitors to examine the role of STAT1 and STAT3 activation. Results: IL-27 increased CXCR3 ligands production in TNF-α-stimulated TR146. Meanwhile, IL-27 suppressed IL-8 and CCL20 production induced by TNF-α. STAT1 phosphorylation level in IL-27 and TNF-α-stimulated TR146 was enhanced in comparison to TNF-α-stimulated TR146. STAT3 phosphorylation level in IL-27-treated TR146 did not change by TNF-α. Both STAT1 inhibitor and STAT3 inhibitor decreased CXCR3 ligands production. STAT1 inhibitor overrode the inhibitory effect of IL-27 on IL-8 and CCL20 production in TNF-α-stimulated TR146. Meanwhile, STAT3 inhibitor did not modulate IL-8 and CCL20 production. Conclusion: IL-27 might control leukocyte migration in periodontal lesion by modulating chemokine production from epithelial cells.

scholarly journals A53 STUDYING THE ROLE OF ASCL2 IN THE ESOPHAGEAL EPITHELIUM USING ORGANOIDS

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 10-11
Author(s):  
M Hamilton ◽  
D Jean ◽  
V Giroux
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Epithelial Cells ◽  
Notch Pathway ◽  
Stem Cell Population ◽  
Esophageal Epithelium ◽  
Ki 67 ◽  
Self Renewal ◽  
Esophageal Epithelial Cells

Abstract Background The esophagus is lined with a stratified squamous epithelium that assure protection against the austere environment found in the esophageal lumen. The maintenance of this epithelium is ensured by a rare population of cells: stem cells. Those cells have increased capacity of self-renewal and multipotency, which is the capacity to give rise to every cell types of a tissue. The marker Krt15 was used to identify the first stem cell population in the esophagus. Krt15+ cells display an extended lifespan and they are radioresistant, multipotent and capable of self-renewal. Moreover, it was observed by RNA sequencing that the expression of the transcription factor ASCL2 is strongly increased in Krt15+ cells compared to Krt15- cells. Interestingly, ASCL2 is necessary to maintain the stemness of Lgr5+ intestinal stem cells. It is also a target of the Wnt/β-catenin pathway. The overall goal of this project is to determine the role of ACSL2 in the maintenance of esophageal stem cells and to identify its binding partners since ASCL2 needs to dimerize to efficiently bind DNA. Aims Confirm that esophageal organoids are adapted to study ASCL2 in the esophagus. Methods Esophageal organoids were established from esophageal epithelial cells from wildtype mice. Following this, organoids were treated with an inhibitor of the Notch pathway (DAPT) to induce hyperplasia or infected with lentiviruses to invalidate Ascl2 (CRISPR/Cas9 approach). Results To validate that Ascl2 plays an important role in esophageal cell proliferation, Notch pathway was inhibited through DAPT treatment in esophageal organoids to induce hyperplasia, which was confirmed by increased number of proliferative cells (Ki-67+). ASCL2 protein expression was also increased in DAPT-treated organoids supporting its role in proliferation and confirming that organoid is a good model to study ASCL2 role in esophageal epithelial cells. In this optic, organoids lines invalidated for Ascl2 (CRISPR/Cas9 approach) were established. Our preliminary results suggest that Ascl2 loss affects cell proliferation and organoid size under normal conditions. Conclusions The expression of ASCL2 correlates with hyperplasia which supports its role in esophageal epithelium homeostasis. Funding Agencies Canada research chair et NSERC

scholarly journals Acidic pH induced NFkB activation and IL‐6 secretion in human esophageal epithelial cells (HET‐1) mediated by PKC

2007 ◽  
Vol 21 (6) ◽  
Author(s):  
Parvaneh Rafiee ◽  
Victoria M Nelson ◽  
Guy Gniotczynski ◽  
Irshad Ali ◽  
David G Binion ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Acidic Ph ◽  
Nfkb Activation ◽  
Esophageal Epithelial Cells

A vH+-ATPase is present in cultured sheep ruminal epithelial cells

2006 ◽  
Vol 291 (6) ◽  
pp. G1171-G1179 ◽  
Author(s):  
Benjamin Etschmann ◽  
Katrin Sophie Heipertz ◽  
Annabelle von der Schulenburg ◽  
Monika Schweigel
Keyword(s):  
Epithelial Cells ◽  
Primary Cultures ◽  
Selective Inhibition ◽  
Protein Band ◽  
Inhibitory Effect ◽  
B Subunit ◽  
Initial Ph ◽  
Acid Load ◽  
Acid Loading

In this study, the existence and functional activity of a vacuolar-type H+-ATPase (vH+-ATPase) was explored in primary cultures of sheep ruminal epithelial cells (REC). The mRNA transcripts of the E and B subunits of vH+-ATPase were detectable in RNA from REC samples by RT-PCR. Immunoblotting of REC protein extractions with antibodies directed against the B subunit of yeast vH+-ATPase revealed a protein band of the expected size (60 kDa). Using the fluorescent indicator BCECF and selective inhibitors (foliomycin, HOE 694, S3226), the contribution of vH+-ATPase and Na+/H+ exchanger (NHE) subtype 1 and 3 activity to the regulation of intracellular pH (pHi) was determined in nominally HCO3−-free, HEPES-buffered NaCl medium containing 20 mM of the short-chain fatty acid butyrate as well as after reduction of the extracellular Cl− concentration ([Cl−]e) from 136 to 36 mM. The initial pHi of REC was 7.4 ± 0.1 in nominally HCO3−-free, HEPES-buffered NaCl medium and 7.0 ± 0.1 after acid loading with butyrate. Selective inhibition of the vH+-ATPase with foliomycin decreased pHi by 0.19 ± 0.03 pH units. On the basis of the observed decreases in pHi resulting from inhibition of vH+-ATPase as well as of subtypes 1 and 3 of NHE, vH+-ATPase activity appears to account for ∼30% of H+ extrusion, whereas the activities of NHE subtypes 3 and 1 account for 20 and 50% of H+ extrusion, respectively. Lowering of [Cl−]e induced a pHi decrease (−0.51 ± 0.03 pH units) and impaired pHi recovery from butyrate-induced acid load. Moreover, reduction of [Cl−]e abolished the inhibitory effect of foliomycin and markedly reduced the HOE 694- and S3226-sensitive components of pHi, indicating a role of Cl− in the function of these H+ extrusion mechanisms. We conclude that a vH+-ATPase is expressed in ovine REC and plays a considerable role in the pHi regulation of these cells.

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