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      Fatty Acid Cytotoxicity to Organ-Cultured Bovine Lenses

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          Abstract

          High sensitivity of cultured bovine and human lens epithelial cells to unsaturated free fatty acids has been reported in previous papers. Here we show that micromolar concentrations of unsaturated free fatty acids also impair lens cells during organ culturing, a system which resembles the in vivo situation much better than cell cultures do. This added weight to the assumption that fatty acid cytotoxicity might be causally related to senile cataract as well as to cataracts associated with systemic diseases.

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          Fatty acid cytotoxicity to human lens epithelial cells.

          Martin IWIG, Dietmar Glaesser, Uwe Fass … (2004)
          Data obtained with the neutral red cytotoxicity assay reveal that human lens epithelial cells in culture are highly sensitive to low micromolar concentrations of unsaturated, cis-configured fatty acids in the following order: arachidonic acid>linolenic acid=linoleic acid=oleic acid, whereas the saturated fatty acids are much less effective. Though the cytotoxic effects of the unsaturated fatty acids could not be discerned from effects of their oxidation products, the fact that oleic acid is equally cytotoxic as linoleic acid or linolenic acid as well as previously reported findings with bovine lens epithelial cells support the idea that the unsaturated fatty acid molecules directly account for the cytotoxicity and not their products of lipid peroxidation. Bleb formation and cell retraction are early morphological signs of fatty acid-induced lens cell damage. These cellular alterations are accompanied by an aggregation of intermediate filaments in a first step, whereas the disorganization of microfilaments occurs at a later time and only at higher fatty acid concentrations. Measurements of protein-, RNA- and DNA-synthesis turned out to be much less sensitive parameters for the fatty acid-induced damage of lens cells. The uptake rate of linoleic acid by human lens cells is relatively high (4.35 fmol sec(-1) per 1000 cells), 30 and 50% higher as compared with diploid human embryonal lung fibroblasts and chemically transformed mouse fibroblasts, respectively. Saturation kinetics in combination with competition between linoleic acid, oleic acid and palmitic acid on one hand and ineffectiveness of trypsin and DIDS treatment on the other hand hint at cytoplasmic fatty acid binding proteins as receptors with high binding affinity (5.55 micromol l(-1), calculated for the linoleic acid-albumin complex) to be involved in the fatty acid uptake in human lens cells. Cellular fatty acid uptake is mainly influenced by the albumin concentrations present in physiological solutions. Albumin determinations in aqueous humor from 177 cataract patients reveal an age-dependent, statistically significant albumin rise with average values below 2 micromol l(-1) up to the age of 40 years to about 4 micromol l(-1) at the age between 80 and 90 years with single values up to 10 micromol l(-1). Using physiological fatty acid mixtures it is demonstrated that fatty acid-induced lens cell damage is strongly increased by elevated albumin concentrations found in aqueous humor of the elderly, who already have cataracts. Free fatty acid induced lens cell damage as a possible cause for age-dependent cataracts as well as a molecular link between systemic diseases such as diabetes and cataract formation is discussed.
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            Linoleic Acid Cytotoxicity to Bovine Lens Epithelial Cells: Influence of Albumin on Linoleic Acid Uptake and Cytotoxicity

            Michael Trimborn, Martin IWIG, Dagobert Glanz … (2000)
            The high cytotoxicity of linoleic acid (LA) to cultured bovine lens epithelial cells is correlated with high uptake rates for the fatty acid (FA). Both, LA uptake and LA cytotoxicity strongly increase with the increasing LA-to-albumin molar ratio in the culture medium. Cellular uptake and cytotoxicity of LA can be competitively inhibited with the noncytotoxic palmitic acid. The findings may be of interest in view of the low albumin concentration in aqueous humor, resulting in extremely low buffering capacities for free FAs including LA, oleic acid and other cytotoxic cis-unsaturated free FAs, which are strongly raised in pathological situations like diabetes mellitus.
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              Fatty acid cytotoxicity to bovine lens epithelial cells: investigations on cell viability, ecto-ATPase, Na(+), K(+)-ATPase and intracellular sodium concentrations.

              N. Nguyen, D Glanz, D Glaesser (2000)
              Unsaturated non-esterified fatty acids have been shown to be cytotoxic in micromolar concentrations to bovine lens epithelial cells, in the following order: arachidonic acid > linoleic acid > oleic acid = linolenic acid. As unsaturated free fatty acids are known to be Na(+), K(+)-ATPase inhibitors, the aim of the study was to investigate whether or not the fatty acid cytotoxicity is correlated with effects on Na(+), K(+)-ATPase activity and function in bovine lens epithelial cells. Furthermore, we also examined the effects of linoleic acid on an ecto-ATPase activity which could be demonstrated on the outside of primarily cultured bovine lens epithelial cells. It has already been shown that 10 micro mol l(-1)linoleic acid was cytotoxic but did not impair the ecto-ATPase activity of intact cells nor the Na(+), K(+)-ATPase in enriched membrane fractions. Na(+), K(+)-ATPase activity was slightly activated with 10 micro mol l(-1)linoleic acid and inhibited by about 50% with 100 micro mol l(-1). Using the sodium-binding benzofuran isophthalate, measurements of intracellular sodium concentrations were carried out. In serum-starved bovine lens epithelial cells the basal [Na(+)](in)was clearly lower than 5 mmol l(-1). When the function of the Na(+), K(+)-ATPase was interrupted by omitting K(+)-ions from the medium, [Na(+)](in)increased at a rate of 0.318 mmol l(-1)min(-1). Linoleic acid intensified that increase strongly in a concentration dependent manner. However, in K(+)-containing medium the linoleic acid-induced increase of [Na(+)](in)was completely prevented. Therefore, the high linoleioc acid cytotoxicity cannot be mediated by linoleic acid effects on Na(+), K(+)-ATPase activity and function in bovine lens epithelial cells.
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                Author and article information

                Journal
                Journal ID (publisher-id): ORE
                Journal ID (nlm-ta): Ophthalmic Res
                Journal ID (doi): 10.1159/issn.0030-3747
                Title: Ophthalmic Research
                Publisher: S. Karger AG
                ISSN (Print): 0030-3747
                ISSN (Electronic): 1423-0259
                Publication date Subscription-year: 2006
                Publication date (Print): February 2006
                Publication date (Electronic): 01 March 2006
                Volume: 38
                Issue: 2
                Pages: 62-65
                Affiliations
                Institute of Physiological Chemistry, Faculty of Medicine, Martin Luther University Halle-Wittenberg, Halle, Germany
                Article
                Publisher ID: 91389 Other ID: Ophthalmic Res 2006;38:62–65
                DOI: 10.1159/000091389
                PubMed ID: 16465068
                SO-VID: 05869d92-4e00-41af-b0c3-9b1d0944a9ab
                Copyright © © 2006 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 : 22 July 2005
                Date accepted : 11 October 2005
                Page count
                Figures: 3, References: 7, Pages: 4
                Categories
                Subject: Short Communication

                ScienceOpen disciplines: Vision sciences,Ophthalmology & Optometry,Pathology
                Keywords: Lenses, bovine,Fatty acid cytotoxicity,Epithelial cells,Organ culture,Fiber cells,Cataract
                Data availability:
                ScienceOpen disciplines: Vision sciences, Ophthalmology & Optometry, Pathology
                Keywords: Lenses, bovine, Fatty acid cytotoxicity, Epithelial cells, Organ culture, Fiber cells, Cataract

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