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Acetate Treatment Increases Fatty Acid Content in LPS-Stimulated BV2 Microglia

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  • Published:
Lipids

Abstract

Acetate supplementation increases plasma acetate, brain acetyl-CoA, histone acetylation, phosphocreatine levels, and is anti-inflammatory in models of neuroinflammation and neuroborreliosis. Although radiolabeled acetate is incorporated into the cellular lipid pools, the effect that acetate supplementation has on lipid deposition has not been quantified. To determine the impact acetate-treatment has on cellular lipid content, we investigated the effect of acetate in the presence of bacterial lipopolysaccharide (LPS) on fatty acid, phospholipid, and cholesterol content in BV2 microglia. We found that 1, 5, and 10 mM of acetate in the presence of LPS increased the total fatty acid content in BV2 cells by 23, 34, and 14 % at 2 h, respectively. Significant increases in individual fatty acids were also observed with all acetate concentrations tested with the greatest increases occurring with 5 mM acetate in the presence of LPS. Treatment with 5 mM acetate in the absence of LPS increased total cholesterol levels by 11 %. However, neither treatment in the absence of LPS significantly altered the content of individual phospholipids or total phospholipid content. To determine the minimum effective concentration of acetate we measured the time- and concentration-dependent changes in histone acetylation using western blot analysis. These studies showed that 5 mM acetate was necessary to induce histone acetylation and at 10 mM acetate, the histone acetylation-state increased as early as 0.5 h following the start of treatment. These data suggest that acetate increases fatty acid content in LPS-stimulated BV2 microglia that is reflected by an increase in fatty acids esterified into membrane phospholipids.

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Abbreviations

ACC:

Acetyl-CoA carboxylase

AMP:

Adenosine monophosphate

ARA:

Arachidonate

ChoGpl:

Choline glycerophospholipid

CerPCho:

Sphingomyelin

DGLA:

Dihomo-γ-linoleate

DHA:

Docosahexaenoate

DMEM/F12:

Dulbecco’s modified eagle medium/nutrient mixture F-12

EtnGpl:

Ethanolamine glycerophospholipid

EDTA:

Ethylenediamine tetraacetic acid

EGTA:

Ethylene glycol tetraacetic acid

EPA:

Eicosapentaenoate

FBS:

Fetal bovine serum

GTA:

Glyceryl triacetate

H3K9:

Histone H3 lysine 9

H4S1/K5/K8/K12:

Histone H4 serine 1, and lysine 5, 8, or 12

HEPES:

2-[4-(2-Hydroxyethyl)piperazine-1-yl]ethanesulfonic acid

HMGCS:

3-Hydroxy-3-methylglutaryl CoA synthase

KH2PO4 :

Potassium phosphate

LNA:

Linoleate

LPS:

Lipopolysaccharide

NaOAc:

Sodium acetate

NaCl:

Sodium chloride

OLA:

Oleate

PAM:

Palmitate

PBS:

Phosphate buffer saline

PtdIns:

Phosphatidylinositol

PtdSer:

Phosphatidylserine

SDS:

Sodium dodecyl sulfate

STA:

Stearate

TTBS:

Tris buffered saline containing Tween 20

TCA:

Tricarboxylic acid

TLC:

Thin layer chromatography

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Acknowledgments

This publication was supported by a University of North Dakota School of Medicine and Health Sciences seed grant and a grant from the NIH/NIGMS (P30GM103329).

Conflict of interest

The authors declare no conflict of interests with the publication of this manuscript.

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Authors and Affiliations

  1. Department of Basic Sciences, School of Medicine and Health Sciences, University of North Dakota, 501 North Columbia Road, Room 3742, Grand Forks, ND, 58203, USA

    Dhaval P. Bhatt & Thad A. Rosenberger

Authors
  1. Dhaval P. Bhatt
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  2. Thad A. Rosenberger
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Correspondence to Thad A. Rosenberger.

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Bhatt, D.P., Rosenberger, T.A. Acetate Treatment Increases Fatty Acid Content in LPS-Stimulated BV2 Microglia. Lipids 49, 621–631 (2014). https://doi.org/10.1007/s11745-014-3911-x

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  • DOI: https://doi.org/10.1007/s11745-014-3911-x

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