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

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

10.21203/rs.3.rs-774286/v1 ◽

2021 ◽

Author(s):

Yuki Kimura ◽

Hideyuki Ohzawa ◽

Hideyo Miyato ◽

Yuki Kaneko ◽

Kazuya Takahashi ◽

...

Keyword(s):

Morphological Changes ◽

Negative Control ◽

Mesothelial Cells ◽

Peritoneal Metastases ◽

Dependent Manner ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

And Migration

Abstract Background: Peritoneal 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). Methods: Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-b1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. Results: HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-b1. TGF-b1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-b1 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. Conclusion: MiR-29b inhibits TGF-b1 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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Role of Peritoneal Mesothelial Cells and Fibroblasts in the Synthesis of Hyaluronan during Peritoneal Dialysis

Peritoneal Dialysis International ◽

10.1177/089686089801800406 ◽

1998 ◽

Vol 18 (4) ◽

pp. 382-386 ◽

Cited By ~ 19

Author(s):

Andrzej Breborowicz ◽

Justyna Wisniewska ◽

Alicja Polubinska ◽

Katarzyna Wieczorowska Tobis ◽

Leo Martis ◽

...

Keyword(s):

Peritoneal Dialysis ◽

Interleukin 1 ◽

Primary Cultures ◽

Mesothelial Cells ◽

Synthesis Rate ◽

Weak Correlation ◽

In Vitro Synthesis ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

Objective To assess the in vitro synthesis rate of hyaluronan (HA) by human peritoneal mesothelial cells and peritoneal fibroblasts in the presence of effluent dialysate from continuous ambulatory peritoneal dialysis (CAPD) patients. Methods We used primary cultures of human peritoneal mesothelial cells and peritoneal fibroblasts from nonuremic patients to study the effect of interleukin-1 β (11–1 β) and pooled effluent dialysate, from noninfected and infected CAPD patients, on the synthesis of HA by the studied cells. We also tested the effect of the exogenous HA on the synthesis rate of that glycosaminoglycan. We studied the correlation between HA concentration in effluent dialysate and the stimulatory effect of that solution on in vitro synthesis of HA by mesothelium. Results Peritoneal fibroblasts produce more HA than mesothelial cells. Noninfected effluent dialysates or dialysates from CAPD patients with peritonitis stimulate synthesis of HA by mesothelial cells and fibroblasts. Interleukin-1 β has a stimulating effect, which was synergistic with effluent dialysates, on the synthesis of HA by mesothelium and peritoneal fibroblasts. A weak correlation was demonstrated between the level of HA in effluent dialysate and the stimulatory effect of that dialysate on in vitro synthesis of HA by mesothelial cells. Conclusions Peritoneal fibroblasts are a more potent source of HA than are mesothelial cells, but probably the latter are the main source of HA in drained dialysate. Although effluent dialysates contain factors that stimulate the production of HA by mesothelium, there is weak correlation between that stimulatory effect and the actual HA concentration in the dialysate, which, in some patients, might suggest low “responsiveness” of the membrane.

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Expression of glucose transporters in human peritoneal mesothelial cells

Kidney International ◽

10.1046/j.1523-1755.1998.00899.x ◽

1998 ◽

Vol 53 (5) ◽

pp. 1278-1287 ◽

Cited By ~ 30

Author(s):

Bernd Schröppel ◽

Michael Fischereder ◽

Patrick Wiese ◽

Stephan Segerer ◽

Stephan Huber ◽

...

Keyword(s):

Glucose Transporters ◽

Mesothelial Cells ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

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

The International Journal of Artificial Organs ◽

10.5301/ijao.2011.8467 ◽

2011 ◽

Vol 34 (6) ◽

pp. 489-494 ◽

Cited By ~ 4

Author(s):

Marta Ciszewicz ◽

George Wu ◽

Paul Tam ◽

Alicja Połubinska ◽

Andrzej Bręborowicz

Keyword(s):

Mesothelial Cells ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

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Catabolism of Newly Synthesized Decorin in vitro by Human Peritoneal Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686080402400205 ◽

2004 ◽

Vol 24 (2) ◽

pp. 147-155 ◽

Cited By ~ 6

Author(s):

Susan Yung ◽

Heinz Hausser ◽

Gareth Thomas ◽

Liliana Schaefer ◽

Hans Kresse ◽

...

Keyword(s):

Dermatan Sulfate ◽

Specific Activity ◽

Cell Layer ◽

Mesothelial Cells ◽

Matrix Assembly ◽

Peritoneal Mesothelial Cells ◽

Intracellular Degradation ◽

Human Peritoneal Mesothelial Cells ◽

Cell Medium

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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3,4-Dideoxyglucosone-3-Ene Induces Apoptosis in Human Peritoneal Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686080902900107 ◽

2009 ◽

Vol 29 (1) ◽

pp. 44-51 ◽

Cited By ~ 15

Author(s):

Duk-Hyun Lee ◽

Soon-Youn Choi ◽

Hye-Myung Ryu ◽

Chan-Duck Kim ◽

Sun-Hee Park ◽

...

Keyword(s):

Flow Cytometry ◽

Caspase 3 ◽

Degradation Products ◽

Treated Group ◽

Mesothelial Cells ◽

Tunel Assay ◽

Dependent Manner ◽

Caspase Inhibitor ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

Objective Glucose degradation products (GDPs) are formed during heat sterilization and storage of peritoneal dialysis (PD) fluids. 3,4-dideoxyglucosone-3-ene (3,4-DGE) has been identified as the most bioreactive GDP. 3,4-DGE induces apoptosis in leukocytes and renal tubular epithelial cells. Our aim was to evaluate the apoptotic effects of 3,4-DGE on human peritoneal mesothelial cells (HPMCs). Methods Primary cultured HPMCs were treated with 25 or 50 μmol/L 3,4-DGE. MTT assay was used to determine cell viability. Apoptosis was measured using TUNEL assay and flow cytometry. Expressions of procaspase-3, Bax, and Bcl-2 were estimated by Western blot. Activity of caspase-3 was measured and the effect of the caspase inhibitor zVAD-fmk (Z-Val-Ala-DL-Asp-fluoromethylketone) was evaluated by TUNEL assay. Results 3,4-DGE treatment accelerated cell death in HPMCs in a dose- and time-dependent manner. Treatment with 3,4-DGE (25 and 50 μmol/L) significantly increased apoptosis compared to control ( p < 0.05 and p < 0.01 respectively) by TUNEL assay. Flow cytometry showed treatment with 50 μmol/L 3,4-DGE significantly increased apoptosis compared to control ( p < 0.05). Decreased expression of procaspase-3 and increased activity of caspase-3 were observed in the presence of 50 μmol/L 3,4-DGE compared to control and 25 μmol/L 3,4-DGE ( p < 0.05). 3,4-DGE-induced HPMC apoptosis was decreased after pretreatment with the pan-caspase inhibitor zVAD-fmk in the 50 μmol/L 3,4-DGE-treated group ( p < 0.001). The ratio of Bcl-2 to Bax expression was decreased in the 25 μmol/L and the 50 μmol/L 3,4-DGE-treated groups compared to control ( p < 0.05). Conclusions 3,4-DGE promotes apoptosis in HPMCs by a caspase-related mechanism.

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Functional Expression of Sodium-Dependent Glucose Transporter in Amelogenesis

Journal of Dental Research ◽

10.1177/0022034520916130 ◽

2020 ◽

Vol 99 (8) ◽

pp. 977-986

Author(s):

H. Ida-Yonemochi ◽

K. Otsu ◽

H. Harada ◽

H. Ohshima

Keyword(s):

Organ Culture ◽

Glucose Uptake ◽

Glucose Transport ◽

Glucose Transporter ◽

Glucose Transporters ◽

Maturation Stage ◽

Sodium Dependent ◽

Tooth Morphogenesis

Glucose is an essential source of energy for mammalian cells and is transported into the cells by glucose transporters. There are 2 types of glucose transporters: one is a passive glucose transporter, GLUT ( SLC2A), and the other is a sodium-dependent active glucose transporter, SGLT ( SLC5A). We previously reported that the expression of GLUTs during tooth development is precisely and spatiotemporally controlled and that the glucose uptake mediated by GLUT1 plays a crucial role in early tooth morphogenesis and tooth size determination. This study aimed to clarify the localization and roles of SGLT1 and SGLT2 in murine ameloblast differentiation by using immunohistochemistry, immunoelectron microscopy, an in vitro tooth organ culture experiment, and in vivo administration of an inhibitor of SGLT1/2, phloridzin. SGLT1, which has high affinity with glucose, was immunolocalized in the early secretory ameloblasts and the ruffle-ended ameloblasts in the maturation stage. However, SGLT2, which has high glucose transport capacity, was observed in the stratum intermedium, papillary layer, and ameloblasts at the maturation stage and colocalized with Na+-K+-ATPase. The inhibition of SGLT1/2 by phloridzin in the tooth germs induced the disturbance of ameloblast differentiation and enamel matrix formation both in vitro (organ culture) and in vivo (mouse model). The expression of SGLT1 and SGLT2 was significantly upregulated in hypoxic conditions in the ameloblast-lineage cells. These findings suggest that the active glucose uptake mediated by SGLT1 and SGLT2 is strictly regulated and dependent on the intra- and extracellular microenvironments during tooth morphogenesis and that the appropriate passive and active glucose transport is an essential event in amelogenesis.

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