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Cytotoxicity of advanced glycation endproducts in human micro- and astroglial cell lines depends on the degree of protein glycation

  • Alzheimer's Disease and Related Disorders - Original Article
  • Published:
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Abstract

Advanced glycation endproducts (AGEs) arise from the reaction of sugars with side chains and the N-terminus of proteins and are thought to be involved in the pathogenesis of several diseases by inducing oxidative stress, inflammation and cell death presumably mediated through activation of the receptor of AGE (RAGE). To address the question whether the cell damaging effect of AGE depends on the degree of its protein glycation, differential modified AGEs derived from incubating human serum albumin with increasing concentrations of methyl glyoxal were tested on cell viability, reactive oxygen species (ROS) formation, intracellular ATP levels, and activation of caspases 3/7 in two human glial cell lines, which were used as a model for human glia cells. All AGEs tested, regardless of their degree of modification, were found to induce ROS formation in both microglial (CHME-5) and astroglial cells (U373 MG), while only highly modified AGEs were able to decrease the cell viability and to induce apoptosis. This indicates that apoptotic events may be involved in the change of physiological parameters.

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Abbreviations

A:

Absorbance

AD:

Alzheimer’s disease

AGEs:

Advanced glycation endproducts

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

CTB:

CellTiter Blue

DMEM:

Dulbecco’s modified Eagle’s medium

FCS:

Fetal calf serum

HBSS:

Hank’s balanced salt solution

HSA:

Human serum albumin

H2DCF-DA:

Dichlorodihydrofluorescein diacetate

MG:

Methyl glyoxal

PB:

Phosphate buffer

PBS:

Phosphate buffered saline

RAGE:

Receptor for AGEs

ROS:

Reactive oxygen species

RT:

Room temperature

TFA:

Trifluoracetoacid

w:

Weeks

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Acknowledgments

This work was supported by the Interdisciplinary Centre of Clinical Research (IZKF) at the Faculty of Medicine of the University of Leipzig (01KS9504, Project N1), the Alzheimer Forschungs Initiative e.V. (Project 01805), the Deutsche Forschungsgemeinschaft (DFG) (Project MU 1011/14-1) and the Studienstiftung des deutschen Volkes (to KB). We thank Prof. Dr. Thomas Henle und Dr. Bernd Weigele, TU Dresden, Germany for providing the IgG antiRAGE antibody, Dr. Carol Lovelidge for revising the manuscript, and special thanks go to Dr. Jovana Gasic-Milenkovic for all the help that she gave us in all AGE related questions as long as we had the pleasure of working together.

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Authors and Affiliations

  1. Interdisciplinary Centre of Clinical Research (IZKF), Faculty of Medicine, University of Leipzig, Inselstr. 22, 04103, Leipzig, Germany

    Katrin Bigl, Annett Schmitt & Sven Rothemund

  2. Institute of Biochemistry, Faculty of Medicine, University of Leipzig, Johannisallee 30, 04103, Leipzig, Germany

    Frank Gaunitz

  3. Comparative Genomics Centre, James Cook University, Molecular Science Bldg. 21, Townsville, QLD, 4811, Australia

    Gerald Münch

  4. Paul-Flechsig-Institute (PFI), Faculty of Medicine, University of Leipzig, Jahnallee 59, 04109, Leipzig, Germany

    Reinhard Schliebs & Thomas Arendt

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  1. Katrin Bigl
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  2. Frank Gaunitz
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Correspondence to Katrin Bigl.

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Bigl, K., Gaunitz, F., Schmitt, A. et al. Cytotoxicity of advanced glycation endproducts in human micro- and astroglial cell lines depends on the degree of protein glycation. J Neural Transm 115, 1545–1556 (2008). https://doi.org/10.1007/s00702-008-0126-4

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  • DOI: https://doi.org/10.1007/s00702-008-0126-4

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