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      Call for Papers: Green Renal Replacement Therapy: Caring for the Environment

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      Effect of Haluronan-Supplemented Dialysate on in vitro Function of Human Peritoneal Mesothelial Cells

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          Abstract

          Background: Addition of hyaluronan (HA) to the dialysis solution has been suggested as a means to protect the peritoneum from injury during peritoneal dialysis (PD). Methods: Concentrations of inflammatory mediators were determined in dialysate samples obtained from PD patients after 6-hour dwells with glucose-based (13.6 g/l) solution containing 0.1 and 0.5 g/l of exogenous high-molecular-weight HA. We additionally evaluated the effect of HA-supplemented dialysate, drained after dwell in PD patients, on function of human peritoneal mesothelial cells (MC) in in vitro culture. Results: Concentration of nitrites was significantly higher in HA 0.5 g/l supplemented dialysate (+43%, p < 0.05) as compared to control. Levels of monocyte chemoattractant protein (MCP-1), soluble intercellular adhesive molecule (s-ICAM), vascular endothelial growth factor (VEGF) and fibronectin were comparable in all the studied groups. However, when MC were exposed in in vitro conditions for 24 h to the studied dialysates, we observed that HA containing fluids inhibited the synthesis of MCP-1, s-ICAM, VEGF and fibronectin in these cells. HA-supplemented dialysate accelerated growth rate of in vitro proliferating MC. Conclusion: High-molecular-weight HA added to the dialysis fluid exerts anti-inflammatory and antifibrotic actions on the in vitro cultured MC and accelerates their growth rate what may be important for peritoneal healing during PD.

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          Hyaluronan Fragments Induce Nitric-oxide Synthase in Murine Macrophages through a Nuclear Factor κB-dependent Mechanism

          Paul Noble, Augustine Choi, Beek Yoke Chin … (1997)
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            A spectrophotometric assay for nitrate using NADPH oxidation by Aspergillus nitrate reductase.

            James M. Gilliam, Stephanie M Sherman, L. Ignarro … (1993)
            An assay based on the oxidation of NADPH during the enzymatic conversion of nitrate to nitrate by Aspergillus nitrate reductase [EC 1.6.6.2] was developed for specific quantification of nitrate. This spectrophotometric method was used to measure nitrate present in human urine, human serum, and tissue culture medium. Used as a kinetic assay, the method exhibited (1) linearity over a range of 1.25 to 40 microM nitrate, (2) an upper sensitivity of 20 microM, (3) a lower sensitivity of 1.25 microM nitrate, and (4) intraday and interday variability ranging from 0.6 to 6.1%. To judge the acceptability of this method as a kinetic assay, we determined the Km for Aspergillus nitrate reductase to be 199 microM. The Km was based on analyzing three separate lots of commercially purified enzyme. Mean nitrate content of eight urine specimens analyzed by this assay (1111 microM) was not significantly different from that determined by a chemiluminescence method (1144 microM). Analysis of serum using the two methods showed mean nitrate concentrations of 23 and 36 microM, respectively. Based on serial dilutions of serum, the lower nitrate content of serum observed with nitrate reductase assay could not be explained by the presence of inhibitors. Rat pulmonary alveolar macrophages were induced to produce nitric oxide which oxidizes to nitrite and nitrate. Nitrite and nitrate present in tissue culture medium of unactivated and activated macrophages were in proportion to total nitrogen oxides (NO(x)) determined by the chemiluminescence method. We conclude that the Aspergillus nitrate reductase assay is an accurate spectrophotometric method for determining nitrate content of human urine and tissue culture supernatants.
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              Stimulation of inducible nitric oxide synthase in rat liver by hyaluronan fragments.

              Paul Chung, D Rockey, Geoffrey Noble … (1997)
              Hepatic injury and chronic wounding are characterized by increased synthesis of extracellular matrix proteins including hyaluronan (HA). Recently, it has been recognized that low-molecular-weight fragments of HA, but not native HA (e.g., high-molecular-weight HA), induce inflammatory gene expression, and activate the transcriptional regulator, nuclear factor kappaB (NF-kappaB). The inducible isoform of nitric oxide synthase (iNOS) is induced by cytokines and/or lipopolysaccharide (LPS) through the NF-kappaB signal transduction pathway. Because of this association, we hypothesized that HA fragments might also stimulate iNOS gene transcription. The aims of this study were therefore to determine whether HA or HA fragments induced iNOS in hepatic cells, and to characterize the signaling pathway. HA fragments (100 microg/mL) markedly stimulated iNOS messenger RNA (mRNA) in endothelial and Kupffer cells, but minimally induced this mRNA in hepatocytes and stellate cells. High-molecular-weight HA (200 microg/mL) had no effect on iNOS mRNA in any cell type. The addition of interferon gamma (IFN-gamma) to HA fragments resulted in stimulation of iNOS mRNA 2-, 3-, 4-, and 10-fold above that for HA fragments alone in hepatocytes, endothelial, Kupffer, and stellate cells, respectively. The combination of HA fragments and LPS did not result in an incremental increase in iNOS mRNA induction. iNOS protein and nitrite levels (used as a measure of NO production and NOS enzymatic activity) paralleled closely iNOS mRNA expression and increased proportionally to HA fragment concentration in a dose-dependent fashion. At 1 hour following stimulation, NF-kappaB DNA binding activity was detected in extracts from Kupffer cells stimulated with HA fragments, but not in those exposed to media alone or to high-molecular-weight HA. Finally, inhibitors of NF-kappaB blocked HA fragment-dependent iNOS mRNA induction in Kupffer and sinusoidal endothelial cells. The data indicate that HA fragments, but not high-molecular-weight HA, induce iNOS in liver, having the greatest effects on endothelial and Kupffer cells. We speculate that HA fragments may be an important stimulus for NO production in various forms of liver disease, particularly as a cofactor with inflammatory cytokines.
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                Author and article information

                Journal
                Journal ID (publisher-id): AJN
                Journal ID (nlm-ta): Am J Nephrol
                Journal ID (doi): 10.1159/issn.0250-8095
                Title: American Journal of Nephrology
                Publisher: S. Karger AG
                ISSN (Print): 0250-8095
                ISSN (Electronic): 1421-9670
                Publication date Subscription-year: 2004
                Publication date (Print): June 2004
                Publication date (Electronic): 06 July 2004
                Volume: 24
                Issue: 3
                Pages: 316-321
                Affiliations
                aDepartment of Pathophysiology, Poznan Medical School, Poznan, Poland; bDivision of Nephrology, Department of Medicine, University of Toronto, Toronto, Ont., Canada; cBaxter Healthcare Corporation, McGraw Park, Ill., USA
                Article
                Publisher ID: 78463 Other ID: Am J Nephrol 2004;24:316–321
                DOI: 10.1159/000078463
                PubMed ID: 15148458
                SO-VID: 20cf0768-f481-47cb-bd72-a7800f269900
                Copyright © © 2004 S. Karger AG, Basel
                License:

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                History
                Date received : 08 March 2004
                Date accepted : 24 March 2004
                Page count
                Figures: 2, Tables: 2, References: 24, Pages: 6
                Categories
                Subject: Original Report: Laboratory Investigation

                ScienceOpen disciplines: Cardiovascular Medicine,Nephrology
                Keywords: Mesothelium,Peritoneal dialysis,Hyaluronan
                Data availability:
                ScienceOpen disciplines: Cardiovascular Medicine, Nephrology
                Keywords: Mesothelium, Peritoneal dialysis, Hyaluronan

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