scholarly journals Effect of Glucose on Intercellular Junctions of Cultured Human Peritoneal Mesothelial Cells

2000 ◽  
Vol 11 (11) ◽  
pp. 1969-1979
Author(s):  
TAKAFUMI ITO ◽  
NORIAKI YORIOKA ◽  
MASAO YAMAMOTO ◽  
KATSUKO KATAOKA ◽  
MICHIO YAMAKIDO
Keyword(s):  
Peritoneal Dialysis ◽  
High Glucose ◽  
Transforming Growth Factor ◽  
Intercellular Junctions ◽  
Neutralizing Antibody ◽  
Transforming Growth Factor Β ◽  
Mesothelial Cells ◽  
Peritoneal Mesothelial Cells ◽  
Peritoneal Dialysis Fluid ◽  
Human Peritoneal Mesothelial Cells

Abstract. During continuous ambulatory peritoneal dialysis, the peritoneum is directly and continuously exposed to unphysiologic peritoneal dialysis fluid; the resulting mesothelial damage has been suggested to cause loss of ultrafiltration and dialysis efficacy. The present study investigated the effect of a high glucose concentration on cultured human peritoneal mesothelial cells to clarify the cause of decreased dialysis efficacy during prolonged peritoneal dialysis. High glucose caused a concentration-dependent decrease in cell proliferation, damage to the intercellular junctions, and excess production of transforming growth factor-β (TGF-β). The levels of intercellular junctional proteins (ZO-1, E-cadherin, and β-catenin) were decreased, and immuno-staining by anti—ZO-1 and anti— β-catenin antibodies became weaker and often discontinuous along the cell contour. Mannitol had similar but weaker effects at the same osmolality, and an anti—TGF-β neutralizing antibody reduced the effects of high glucose. Therefore, these effects were induced not only by glucose itself but also by hyperosmolality and by a glucose-induced increase of TGF-β. These findings suggest that the peritoneal mesothelium is damaged by prolonged peritoneal dialysis using high glucose dialysate and that impairment of the intercellular junctions of peritoneal mesothelial cells by high glucose dialysate induces peritoneal hyperpermeability and a progressive reduction in dialysis efficacy.

Effects of Peritoneal Dialysis Solutions on the Secretion of Growth Factors and Extracellular Matrix Proteins by Human Peritoneal Mesothelial Cells

2002 ◽  
Vol 22 (2) ◽  
pp. 171-177 ◽  
Author(s):  
Hunjoo Ha ◽  
Mi Kyung Cha ◽  
Hoo Nam Choi ◽  
Hi Bahl Lee
Keyword(s):  
Peritoneal Dialysis ◽  
Growth Factor ◽  
High Glucose ◽  
Transforming Growth Factor ◽  
Mesothelial Cells ◽  
Dependent Manner ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells ◽  
Dialysis Solutions ◽  
Peritoneal Dialysis Solutions

♦ Objective To compare the effects of different peritoneal dialysis solutions (PDS) on secretion of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGFβ1), procollagen I C-terminal peptide (PICP), procollagen III N-terminal peptide (PIIINP), and fibronectin by cultured human peritoneal mesothelial cells (HPMC). ♦ Design Using M199 culture medium as control, commercial PDS containing 1.5% or 4.25% glucose and 40 mmol/L lactate [Dianeal 1.5 (D 1.5) and Dianeal 4.25 (D 4.25), respectively; Baxter Healthcare, Deerfield, Illinois, USA]; PDS containing 1.5% or 4.25% glucose with 25 mmol/L bicarbonate and 15 mmol/L lactate [Physioneal 1.5 (P 1.5) and Physioneal 4.25 (P 4.25), respectively; Baxter]; and PDS containing 7.5% icodextrin [Extraneal (E); Baxter] were tested. Growth-arrested and synchronized HPMC were continuously stimulated for 48 hours by test PDS diluted twofold with M199, TGFβ1 1 ng/mL, or different concentrations of icodextrin. VEGF, TGFβ1, and fibronectin secreted into the media were analyzed by ELISA, and PICP and PIIINP by radioimmunoassay. ♦ Results Dianeal 1.5, D 4.25, and P 4.25, but not P 1.5 and E, significantly increased VEGF secretion compared with control M199. D 4.25- and P 4.25-induced VEGF secretion was significantly higher than induction by D 1.5 and P 1.5, respectively, suggesting that high glucose may be involved in the induction of VEGF. Physioneal 1.5- and P 4.25-induced VEGF secretion was significantly lower than induction by D 1.5 and D 4.25, respectively, suggesting a role for glucose degradation products (GDP) in VEGF production. TGFβ1 secretion was significantly increased by D 4.25 and E. Icodextrin increased TGFβ1 secretion in a dose-dependent manner. All PDS tested significantly increased secretion of PIIINP compared with control. D 1.5- and D 4.25-induced PIIINP secretion was significantly higher than P 1.5, P 4.25, and E. Physioneal 4.25-induced PIIINP secretion was significantly higher than P 1.5, again implicating high glucose and GDP in PIIINP secretion by HPMC. There was no significant increase in PICP or fibronectin secretion using any of the PDS tested. Addition of TGFβ1 1 ng/mL into M199 control significantly increased VEGF, PICP, PIIINP, and fibronectin secretion by HPMC. ♦ Conclusions The present study provides direct evidence that HPMC can secrete VEGF, TGFβ1, and PIIINP in response to PDS, and that HPMC may be actively involved in the development and progression of the peritoneal membrane hyperpermeability and fibrosis observed in long-term PD patients. This study also suggests that both high glucose and GDP in PDS may play important roles in inducing VEGF and PIIINP production/secretion by HPMC.

Peritoneal Dialysis Fluid Induces P38-Dependent Inflammation in Human Mesothelial Cells

2011 ◽  
Vol 31 (3) ◽  
pp. 332-339 ◽  
Author(s):  
Andrea Riesenhuber ◽  
Klaus Kratochwill ◽  
Thorsten O. Bender ◽  
Regina Vargha ◽  
David C. Kasper ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Stress Response ◽  
Stress Responses ◽  
Mesothelial Cells ◽  
Hsp70 Expression ◽  
Peritoneal Mesothelial Cells ◽  
P38 Inhibitor ◽  
Peritoneal Dialysis Fluid ◽  
Human Peritoneal Mesothelial Cells ◽  
Ldh Release

BackgroundNoninfectious upregulation of proinflammatory pathways in mesothelial cells may represent an integral part of their stress response upon exposure to peritoneal dialysis fluids (PDF).ObjectiveThe aim of this study was to evaluate the role of the stress-inducible mitogen-activated protein kinase (MAPK) p38 in regulation of inflammatory and stress responses in mesothelial cells following in vitro exposure to PDF.Materials and MethodsHuman peritoneal mesothelial cells were exposed to Dianeal PD4 or Physioneal (Baxter AG, Vienna, Austria) containing 1.36% glucose and then allowed to recover. Phosphorylation of p38, induction of heat shock protein-70 (HSP70), release of lactate dehydrogenase (LDH), secretion of interleukin (IL)-8, gene transcription, and mRNA stability were assessed with and without the MAPK p38 inhibitor SB203580.ResultsExposure to Dianeal resulted in phosphorylation of p38 within 30 minutes (309% of control, p < 0.05) and increased IL-8 release (370% of control, p < 0.05), HSP70 expression (151% of control, p < 0.05), and LDH release (180% of control, p < 0.05). Exposure to Physioneal resulted in attenuated changes in IL-8, HSP70, and LDH. Addition of the p38 inhibitor SB203580 to Dianeal resulted in dampened IL-8 release (-55%; p < 0.05) and basal HSP70 expression, and unchanged LDH release. Effects of p38 on IL-8 were at transcriptional, posttranscriptional, and translational levels.ConclusionThese data confirm concordant p38-dependent upregulation of IL-8 and HSP70 following exposure to bioincompatible PDF. The MAPK p38 pathway therefore links proinflammatory processes and the cellular stress response in human peritoneal mesothelial cells.

Emodin Ameliorates Glucose-Induced Morphologic Abnormalities and Synthesis of Transforming Growth Factor β1 and Fibronectin by Human Peritoneal Mesothelial Cells

2001 ◽  
Vol 21 (3_suppl) ◽  
pp. 41-47 ◽  
Author(s):  
Susan Yung ◽  
Zhi-Hong Liu ◽  
Kar-Neng Lai ◽  
Lei-Shi Li ◽  
Tak-Mao Chan
Keyword(s):  
Growth Factor ◽  
High Glucose ◽  
Glucose Concentration ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Mesothelial Cells ◽  
High Glucose Concentration ◽  
Matrix Synthesis ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells

♦ Objective Excessive synthesis and deposition of matrix proteins by peritoneal mesothelial cells can lead to structural and functional changes in the peritoneal membrane, jeopardizing the long-term efficacy of peritoneal dialysis (PD). Prolonged exposure to high glucose concentrations in PD fluid has been implicated as a major stimulus to matrix accumulation, through the induction of transforming growth factor β1 (TGFβ 1). This study investigated the effect of emodin (3-methyl-1,6,8-trihydroxyanthraquinone) on TGFβ 1 and fibronectin (FN) synthesis in human peritoneal mesothelial cells (HPMCs) under high glucose concentration. ♦ Design The HPMCs were preconditioned in either 5 mmol/L or 30 mmol/L d-glucose for 2 weeks prior to the addition of emodin. Cell viability was assessed by MTT assay and lactate dehydrogenase (LDH) release. Morphology of HPMCs was studied by phase-contrast microscopy. Modulation of TGFβ 1 and FN synthesis at transcription and translation were investigated by reverse transcriptase polymerase chain reaction (RT-PCR), ELISA, and Western blot analysis. ♦ Results When cultured under 30 mmol/L d-glucose, HPMCs demonstrated increased cell volume, multi-nucleation, and denudation of the monolayer, as compared with cells cultured under a physiologic (5 mmol/L) glucose concentration. High glucose concentration induced TGFβ 1 synthesis by HPMCs (217.17 ± 14.88 pg/mL at 5 mmol/L d-glucose vs 370.33 ± 20.67 pg/mL at 30 mmol/L d-glucose, p < 0.0001), and FN synthesis was induced at transcription and translation. Mannitol at 30 mmol/L did not affect HPMC morphology; matrix synthesis was also unaltered. Administration of emodin together with 30 mmol/L d-glucose resulted in amelioration of cell enlargement and exfoliation, and abrogation of TGFβ 1 induction (370.33 ± 20.67 pg/mL for 30 mmol/L d-glucose alone vs 260.50 ± 17.89 pg/mL for 30 mmol/L d-glucose + emodin, p < 0.0001). Synthesis of FN induced by high glucose was also reduced by 40% in the presence of emodin. ♦ Conclusions These findings provide the first evidence that emodin can ameliorate high glucose–induced matrix synthesis in HPMCs by suppression of TGFβ 1. Emodin may thus be useful in preserving peritoneal integrity in PD.

scholarly journals Polyphenols Attenuate Highly-Glycosylated Haemoglobin-Induced Damage in Human Peritoneal Mesothelial Cells

Antioxidants ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 572
Author(s):  
Carolina Sánchez-Rodríguez ◽  
Concepción Peiró ◽  
Leocadio Rodríguez-Mañas ◽  
Julián Nevado
Keyword(s):  
Oxidative Stress ◽  
Peritoneal Dialysis ◽  
P53 Protein ◽  
Mesothelial Cells ◽  
Protein Glycation ◽  
Membrane Function ◽  
Peritoneal Mesothelial Cells ◽  
Dietary Polyphenols ◽  
Peritoneal Dialysis Fluid ◽  
Human Peritoneal Mesothelial Cells

We investigated the cytoprotective role of the dietary polyphenols on putative damage induced by Amadori adducts in Human Peritoneal Mesothelial Cells (HPMCs). Increased accumulation of early products of non-enzymatic protein glycation—Amadori adducts—in the peritoneal dialysis fluid due to their high glucose, induces severe damage in mesothelial cells during peritoneal dialysis. Dietary polyphenols reportedly have numerous health benefits in various diseases and have been used as an efficient antioxidant in the context of several oxidative stress-related pathologies. HPMCs isolated from different patients were exposed to Amadori adducts (highly glycated haemoglobin, at physiological concentrations), and subsequently treated with several polyphenols, mostly presented in our Mediterranean diet. We studied several Amadori-induced effects in pro-apoptotic and oxidative stress markers, as well as the expression of several pro-inflammatory genes (nuclear factor-kappaB, NF-kB; inducible Nitric Oxide synthetase, iNOS), different caspase-activities, level of P53 protein or production of different reactive oxygen species in the presence of different polyphenols. In fact, cytoprotective agents such as dietary polyphenols may represent an alternate approach to protect mesothelial cells from the cytotoxicity of Amadori adducts. The interference with the Amadori adducts-triggered mechanisms could represent a therapeutic tool to reduce complications associated with peritoneal dialysis in the peritoneum, helping to maintain peritoneal membrane function longer in patients undergoing peritoneal dialysis.

High Glucose Solution and Spent Dialysate Stimulate the Synthesis of Transforming Growth Factor-β1 of Human Peritoneal Mesothelial Cells: Effect of Cytokine Costimulation

1999 ◽  
Vol 19 (3) ◽  
pp. 221-230 ◽  
Author(s):  
Duk-Hee Kang ◽  
Young-Sook Hong ◽  
Hyun Joung Lim ◽  
Jin-Hee Choi ◽  
Dae-Suk Han ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Protein Secretion ◽  
High Glucose ◽  
Transforming Growth Factor ◽  
Glucose Solution ◽  
Mesothelial Cells ◽  
Peritoneal Dialysate ◽  
Peritoneal Mesothelial Cells ◽  
Spent Dialysate ◽  
Human Peritoneal Mesothelial Cells

Objective To investigate the effect of high glucose and spent peritoneal dialysate on the transforming growth factor-β1 (TGFβ1) synthesis of cultured human peritoneal mesothelial cells (HPMCs) and to examine the effect of costimulation with high glucose or spent dialysate, and cytokines, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNFα) on TGFβ1 synthesis of HPMCs. Design HPMCs were exposed to different concentrations of glucose (30, 60, and 90 mmol/L) or spent peritoneal dialysate for 48 hours in the absence or presence of IL-1β (1 ng/mL) and TNFα (1 ng/mL). TGFβ1 mRNA expression was assessed by Northern blot analysis and TGFβ1 protein release by Western blot analysis and enzymelinked immunosorbent assay (ELISA). Results Exposure of HPMCs to high glucose conditions (30, 60, and 90 mmol/L of D-glucose) induced 2.3-, 3.6-, and 4.0-fold increases in TGFβ1 mRNA expression of HPMC with enhanced TGFβ1 protein synthesis and secretion into the media, whereas there were no significant changes in TGFβ1 synthesis with equimolar concentrations of D-mannitol. Incubation with spent dialysate also significantly increased TGFβ1 mRNA expression and protein secretion compared to control media ( p < 0.05). Stimulation with IL-1β (1 ng/mL) or TNFα (1 ng/mL) resulted in a significant increase in TGFβ1 mRNA expression after 48 hours: 2.7 and 2.1 times the control level, respectively. However, TNFα-induced increase in TGFβ1 mRNA expression was not translated into TGFβ1 protein secretion, while IL-1β stimulation induced a significant increase in TGFβ1 protein secretion as well as TGFβ1 mRNA expression. Combined stimulation by high glucose or spent dialysate, together with IL-1β or TNFα, showed a greater increase in TGFβ1 mRNA expression and protein secretion compared to stimulation by high glucose or spent dialysate alone. Conclusion Our results clearly show that high glucose solution and spent dialysate themselves might be sufficient to stimulate the production of TGFβ1 by peritoneal mesothelial cells. In peritoneal dialysis patients, this state of chronic induction of TGFβ1 is further exacerbated in the presence of peritonitis because of the stimulatory effect of proinflammatory cytokines, resulting in augmented TGFβ1 synthesis, thus promoting peritoneal fibrosis.

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