Glucose but not N-Acetylglucosamine Accelerates in vitro Senescence of Human Peritoneal Mesothelial Cells

Objective Previous studies have shown that decorin and biglycan account for over 70% of the proteoglycans (PGs) synthesized by human peritoneal mesothelial cells (HPMCs). Since these PGs are involved in the control of cell growth, cell differentiation, and matrix assembly, we investigated their turnover in cultured HPMCs. Methods Confluent HPMCs were metabolically labeled with [35S]-sulfate and the labeled products isolated from the cell medium and the cell layer characterized by sensitivity to bacterial eliminases. Experiments were undertaken with exogenous labeled decorin, and its metabolic state was studied. Results In a 24-hour labeling period, 75% of the newly synthesized chondroitin sulfate/dermatan sulfate (CS/DS) PGs appeared in the culture medium, the majority of which (90%) was decorin. In the cell layer, protein-free glycosaminoglycan (GAG) chains accounted for 21% of the total CS/DS at 24 hours and exhibited constant specific activity at 12 – 16 hours. The latter material was turned over with a half-life of approximately 2.5 hours. Exogenous decorin underwent receptor-mediated endocytosis and subsequent intracellular degradation. Uptake but not degradation could be inhibited by heparin. Conclusions HPMCs are distinguished by a rapid turnover of decorin. A characteristic metabolic feature is the existence of a large intracellular pool of protein-free DS-GAGs. Understanding the control of decorin turnover in HPMCs might lead to delineation of its potential role in both the physiology and pathophysiology of the membrane in PD patients.

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Expression of aquaporin-3 in human peritoneal mesothelial cells and its up-regulation by glucose in vitro

Kidney International ◽

10.1111/j.1523-1755.2002.kid564.x ◽

2002 ◽

Vol 62 (4) ◽

pp. 1431-1439 ◽

Cited By ~ 10

Author(s):

Kar Neng Lai ◽

Joseph C.K. Leung ◽

Loretta Y.Y. Chan ◽

Sydney Tang ◽

Fu Keung Li ◽

...

Keyword(s):

Mesothelial Cells ◽

Aquaporin 3 ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

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Mesothelial Cell Adherence to Vascular Prostheses and Their Subsequent Growth In Vitro

Cell Transplantation ◽

10.1177/096368979400300107 ◽

1994 ◽

Vol 3 (1) ◽

pp. 41-48 ◽

Cited By ~ 3

Author(s):

Apollo Pronk ◽

Arthur A.G.M. Hoynck Van Papendrecht ◽

Piet Leguit ◽

Henri A. Verbrugh ◽

Roel P.A.J. Verkooyen ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Attachment ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Subsequent Growth ◽

Vascular Prostheses ◽

Peritoneal Mesothelial Cells ◽

Viable Cells ◽

Human Peritoneal Mesothelial Cells

Cell seeding may decrease the thrombogenicity of implanted vascular grafts, but its application is hampered by the limited availability of autologous endothelial cells. Human peritoneal mesothelial cells have blood flow supporting qualities and are readily available. This study investigated the adherence of mesothelial cells to vascular prostheses and their subsequent growth in vitro. Circular pieces of various vascular prosthetic materials were seeded with 51Chromium-labeled mesothelial and endothelial cells and left for either 5, 15, 30, 60, and 120 minutes. The unattached cells were removed and the degree of cell attachment was measured. The number of mesothelial cells to Dacron increased during the first 60 min up to 35.2 % of the seeded inoculum whereafter a plateau was reached. Scanning electron microscopy showed spreaded mesothelial cells adherent to the Dacron fibers. A significant increase in adherence was observed after preincubation of Dacron with 10 μg/mL fibronectin, but no improvement was found after preincubation with human serum albumin or gelatin. Mesothelial cells adhered better to Gelcoated than to Gelsealed or plain Dacron. The adherence of mesothelial cells to ePTFE (Teflon) was significantly poorer. No significant differences in adherence were found between mesothelial and endothelial cells. Mesothelial cell growth on Dacron resulted in a modest increase in the number of viable cells during 27 days, which implies biocompatibility of Dacron and mesothelial cells in vitro.

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miRNA589 Regulates Epithelial-Mesenchymal Transition in Human Peritoneal Mesothelial Cells

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/673096 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6 ◽

Cited By ~ 15

Author(s):

Ke Zhang ◽

Hao Zhang ◽

Xun Zhou ◽

Wen-bin Tang ◽

Li Xiao ◽

...

Keyword(s):

Epithelial Mesenchymal Transition ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Control Group ◽

Peritoneal Fibrosis ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

E Cadherin

Background. microRNA (miRNA, miR) are thought to interact with multiple mRNAs which are involved in the EMT process. But the role of miRNAs in peritoneal fibrosis has remained unknown.Objective. To determine if miRNA589 regulates the EMT induced by TGFβ1 in human peritoneal mesothelial cell line (HMrSV5 cells).Methods. 1. Level of miR589 was detected in both human peritoneal mesothelial cells (HPMCs) isolated from continuous ambulatory peritoneal dialysis (CAPD) patients’ effluent and HMrSV5 cells treated with or without TGFβ1. 2. HMrSV5 cells were divided into three groups: control group, TGFβ1 group, and pre-miR-589+TGFβ1 group. The level of miRNA589 was determined by realtime PCR. The expressions of ZO-1, vimentin, and E-cadherin in HPMCs were detected, respectively.Results. Decreased level of miRNA589 was obtained in either HPMCs of long-term CAPD patients or HMrSV5 cells treated with TGFβ1. In vitro, TGFβ1 led to upregulation of vimentin and downregulation of ZO-1 as well as E-cadherin in HMrSV5 cells, which suggested EMT, was induced. The changes were accompanied with notably decreased level of miRNA589 in HMrSV5 cells treated with TGFβ1. Overexpression of miRNA589 by transfection with pre-miRNA589 partially reversed these EMT changes.Conclusion. miRNA589 mediates TGFβ1 induced EMT in human peritoneal mesothelial cells.

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Interleukin-1β Stimulates Glucose Uptake of Human Peritoneal Mesothelial Cells In Vitro

Peritoneal Dialysis International ◽

10.1177/089686089601601s09 ◽

1996 ◽

Vol 16 (1_suppl) ◽

pp. 58-60 ◽

Cited By ~ 5

Author(s):

Michael Kruse ◽

Arezki Mahiout ◽

Volker Kliem ◽

Peter Kurz ◽

Karl-Martin Koch ◽

...

Keyword(s):

Glucose Uptake ◽

Glucose Transporter ◽

Cytochalasin B ◽

Interleukin 1 ◽

Glucose Transporters ◽

Mesothelial Cells ◽

Dependent Manner ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

To investigate whether the glucose uptake (GU) of human peritoneal mesothelial cells (HPMC) is mediated by glucose transporters and whether this uptake is influenced by interleukin 1–β (IL-1β), we measured 2-deoxy-(3H)-GU of HPMC in vitro, after exposing the cells for different times (two and 12 hours) to increasing concentrations (0.1, 1.0, and 2.0 ng/mL) of IL-1 β. To exclude a noncarrier-mediated transport, GU was also tested in the presence of cytochalasin B. All experiments were performed in triplicate in the cells of two donors. Cytochalasin B inhibits GU of HPMC almost completely. GU of HPMC is not stimulated by insulin. GU is stimulated by IL-1 β in a dose-dependent manner. These data indicate a GU of HPMC, which is mediated by a glucose transporter and stimulated by IL-1 β. The increased uptake of glucose from the dialysate In patients with peritonitis may be mediated by a (cytokineinduced) increased activity of HPMC glucose transporters.

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Glycerol Toxicity for Human Peritoneal Mesothelial Cells in Culture: Comparison with Glucose

The International Journal of Artificial Organs ◽

10.1177/039139889401700502 ◽

1994 ◽

Vol 17 (5) ◽

pp. 252-260 ◽

Cited By ~ 14

Author(s):

J. Witowski ◽

J. Knapowski

Keyword(s):

Mesothelial Cell ◽

Cellular Protein ◽

Mesothelial Cells ◽

Dependent Manner ◽

Peritoneal Mesothelial Cells ◽

Proline Incorporation ◽

Human Peritoneal Mesothelial Cells ◽

Dose Dependent Decrease ◽

Dose Dependent

Glycerol has been proposed as a substitute osmotic agent for glucose in peritoneal dialysis fluids. We have compared the effect of glycerol and glucose on the function of human peritoneal mesothelial cells (HPMC) in vitro. The viability of HPMC was not affected by glycerol (up to 250 mM), whereas it was reduced by glucose in a time- and dose-dependent manner, as assessed by the LDH release. Although the incubation of HPMC with glycerol induced a dose-dependent decrease in HPMC proliferation, the effect was significantly less inhibitory than that produced by glucose. In HPMC treated with 90 mM of glycerol or glucose the incorporation of [3H]-thymidine had reached 79.0±19.3% and 55.3+4.0% of the control (p<0.05 and p<0.01), respectively. As measured by the [methyl-14C]-choline incorporation, the intracellular amount of newly synthesized phospholipids was reduced from (cpm/μg cellular protein) 147±58 in control HPMC to 59+15 in cells exposed to 90 mM of glucose (p<0.01), but not affected by glycerol (163±65). On the other hand, both glycerol and glucose (90 mM) decreased the synthesis of proteins (as assessed by the [3H]-proline incorporation) and interfered with potassium (86Rb) transport mechanisms in HPMC. Our data suggest that there exist some possibly advantageous aspects of glycerol as far as mesothelial cell biocompatibility profile is concerned.

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In Vitro Cytotoxicity of Amino Acids and Their Glycyl-Dipeptide Forms to Human Peritoneal Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686089401400219 ◽

1994 ◽

Vol 14 (2) ◽

pp. 182-184 ◽

Cited By ~ 1

Author(s):

Andrzej Breborowicz ◽

Janusz Witowski ◽

Jan Knapowski ◽

George D. Oreopoulos ◽

Elias V. Balaskas ◽

...

Keyword(s):

Amino Acids ◽

Mesothelial Cells ◽

In Vitro Cytotoxicity ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Glycyl Dipeptide

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Differential Regulation of Chemokine Production in Human Peritoneal Mesothelial Cells: IFN-γ Controls Neutrophil Migration Across the Mesothelium In Vitro and In Vivo

The Journal of Immunology ◽

10.4049/jimmunol.167.2.1028 ◽

2001 ◽

Vol 167 (2) ◽

pp. 1028-1038 ◽

Cited By ~ 51

Author(s):

Rachel L. Robson ◽

Rachel M. McLoughlin ◽

Janusz Witowski ◽

Pius Loetscher ◽

Thomas S. Wilkinson ◽

...

Keyword(s):

Neutrophil Migration ◽

Differential Regulation ◽

Mesothelial Cells ◽

Chemokine Production ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Ifn Γ

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MiR-29b may suppresses peritoneal metastases through inhibition of the mesothelial–mesenchymal transition (MMT) of human peritoneal mesothelial cells

Scientific Reports ◽

10.1038/s41598-021-04065-2 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Yuki Kimura ◽

Hideyuki Ohzawa ◽

Hideyo Miyato ◽

Yuki Kaneko ◽

Akira Saito ◽

...

Keyword(s):

Morphological Changes ◽

Negative Control ◽

Mesothelial Cells ◽

Peritoneal Metastases ◽

Dependent Manner ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Tgf Β1

AbstractPeritoneal dissemination is a major metastatic pathway for gastrointestinal and ovarian malignancies. The miR-29b family is downregulated in peritoneal fluids in patients with peritoneal metastases (PM). We examined the effect of miR-29b on mesothelial cells (MC) which play critical a role in the development of PM through mesothelial-mesenchymal transition (MMT). Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-β1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-β1. TGF-β1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-β1 also enhanced proliferation and migration of HPMC as well as adhesion of tumor cells in a fibronectin dependent manner. However, all events were strongly abrogated by simultaneous transfection of miR-29b. MiR-29b inhibits TGF-β1 induced MMT and replacement of miR-29b in the peritoneal cavity might be effective to prevent development of PM partly through the effects on MC.

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