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Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia

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An Erratum to this article was published on 04 February 2017

Abstract

Recent evidence suggests that metformin shows beneficial effects in experimental models of neuroinflammatory diseases. The aim of the present study was to determine the effect of metformin on phagocytosis and acidification of lysosomal/endosomal compartments in rat primary microglia in the presence of lipopolysaccharide (LPS) and/or beta-peptides (25–35), (1–40), and (1–42). Metformin increased the phagocytosis of fluorescent microspheres in the presence or absence of all the beta-peptides. However, the drug had no effect on the phagocytosis in LPS-stimulated microglia regardless of the presence of all the beta-peptides. Metformin acidified the lysosomal/endosomal compartments in the presence or absence of the beta-peptide 1–40 in both resting and activated microglia. To elucidate the mechanism of metformin action, we used 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside as an activator of adenosine monophosphate-activated protein kinase (AMPK) and compound C as a confirmed pharmacological inhibitor of AMPK. We have shown that metformin increased AMPK activity in microglial cells and that all observed effects are AMPK-dependent because the pretreatment of microglia with compound C reversed the effects of the drug. Since degradation of proteins in lysosomal/endosomal compartments depends largely on their phagocytosis and acidification, metformin may be beneficial in proteinopathies affecting the brain.

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Abbreviations

AICAR:

5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside

AMPK:

Adenosine monophosphate-activated protein kinase

BrdU:

5-Bromo-2′-deoxyuridine

DMEM:

Dulbecco's modified Eagle's medium

FBS:

Fetal bovine serum

GFAP:

Glial fibrillary acidic protein

IOD:

Integrated optical density

LPS:

Lipopolysaccharide

MAP-2:

Microtubule associating protein-2

MTT:

3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolinum bromide

RCA-1:

Ricinus Communis Agglutinin-1

RPMI-1640 Medium:

Roswell Park Memorial Institute-1640 Medium

TBST:

Tris-buffered saline Tween 20

SD:

Standard deviation

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Acknowledgements

The authors are thankful to Mrs. Jaroslawa Sprada, Mrs. Halina Klimas, and Mrs. Anna Bielecka for their excellent technical support. This work was supported by a research grant KNW-2-092/09 from Medical University of Silesia, Katowice, Poland. None of the authors has any conflict of interest. The study was approved by the Ethical Committee of the Medical University of Silesia. The experiments comply with the current law of Poland.

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Authors and Affiliations

  1. Department of Clinical Pharmacology, Medical University of Silesia, Medyków 18, 40-752, Katowice, Poland

    Krzysztof Łabuzek, Sebastian Liber & Bogusław Okopień

  2. Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752, Katowice, Poland

    Bożena Gabryel

  3. Department of Biophysics, School of Pharmacy, Medical University of Silesia, Jedności 8, 41-200, Sosnowiec, Poland

    Jakub Adamczyk

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  1. Krzysztof Łabuzek
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Correspondence to Krzysztof Łabuzek.

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An erratum to this article is available at http://dx.doi.org/10.1007/s00210-017-1350-y.

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Łabuzek, K., Liber, S., Gabryel, B. et al. Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia. Naunyn-Schmied Arch Pharmacol 381, 171–186 (2010). https://doi.org/10.1007/s00210-009-0477-x

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