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      Effect of Insulin on Hydrogen Peroxide Production by Human Polymorphonuclear Leukocytes

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          Abstract

          This study evaluated the effect of insulin on the respiratory burst of human polymorphonuclear leukocytes (PMNLs) and the signalling pathways involved in this process, especially the involvement of protein kinase C (PKC). Isolated human PMNLs from healthy volunteers were incubated with different concentrations of insulin (10<sup>-10</sup>-10<sup>-7</sup> mol/l) and for different durations of incubation (5-90 min). The intracellular production of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) was detected employing a previously validated flow cytometric assay using 2’,7’-dichlorofluorescein-diacetate (DCFH-DA) as a marker for H<sub>2</sub>O<sub>2 </sub>production. Specificity of insulin action was verified using an insulin antagonist (the monoclonal antibody MA-10). To identify the signalling pathway involved, we used: (a) monoclonal antibody MA-5, directed against the α-subunit of the insulin receptor, that partially mimics insulin without activating tyrosine kinase; (b) H7, an inhibitor of PKC involved in O<sub>2</sub><sup>-</sup> production in PMNLs, and (c) phorbol myristate acetate (PMA) that binds and stimulates PKC. Insulin caused a dose- and time-dependent stimulation of H<sub>2</sub>O<sub>2</sub> release by human PMNLs. The effect of insulin was blocked by MA-10. The actions of insulin and PMA on H<sub>2</sub>O<sub>2</sub> release were additive, whereas the actions of MA-5 and PMA were not. H7 partially inhibited the H<sub>2</sub>O<sub>2</sub> production stimulated by insulin and completely inhibited MA-5 action. We conclude that insulin stimulates, in a dose- and time-related manner, the respiratory burst of human PMNLs. PKC activation can only partially account for the intracellular mechanisms involved in this process.

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          Author and article information

          Journal
          HRE
          Horm Res Paediatr
          10.1159/issn.1663-2818
          Hormone Research in Paediatrics
          S. Karger AG
          1663-2818
          1663-2826
          1995
          1995
          05 December 2008
          : 43
          : 6
          : 286-293
          Affiliations
          Departments of aPediatrics and bInternal Medicine, Tor Vergata University, Rome, Italy
          Article
          184313 Horm Res 1995;43:286–293
          10.1159/000184313
          7607614
          © 1995 S. Karger AG, Basel

          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.

          Page count
          Pages: 8
          Categories
          Original Paper

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