Lipids

, Volume 49, Issue 7, pp 621–631

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

Original Article

DOI: 10.1007/s11745-014-3911-x

Cite this article as:
Bhatt, D.P. & Rosenberger, T.A. Lipids (2014) 49: 621. doi:10.1007/s11745-014-3911-x

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.

Keywords

Acetate Cholesterol Fatty acid Histone acetylation Lipids Phospholipid Microglia 

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

Supplementary material

11745_2014_3911_MOESM1_ESM.docx (24 kb)
Supplementary material 1 (DOCX 23 kb)

Copyright information

© AOCS 2014

Authors and Affiliations

  1. 1.Department of Basic Sciences, School of Medicine and Health SciencesUniversity of North DakotaGrand ForksUSA