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Characterization of apoA-I-dependent lipid efflux from adipocytes and role of ABCA1

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Abstract

Adipose tissue is a major reservoir of cholesterol and, as such, it may play a significant role in cholesterol homeostasis. The aims of this study were to obtain a quantitative characterization of apolipoprotein A-I (apoA-I)-dependent lipid efflux from adipocytes and examine the role of ATP-binding cassette transporter A1 (ABCA1) in this process. The rates of apoA-I-induced cholesterol and phospholipid efflux were determined and normalized by cellular protein or ABCA1 levels. In order to allow a comparative analysis, parallel experiments were also performed in macrophages. These studies showed that apoA-I induces cholesterol efflux from adipocytes at similar rates as from macrophages. Enhancement of the expression of ABCA1 increased the rates of cholesterol efflux from both adipocytes and macrophages. The results also suggested that a non-ABCA1-dependent mechanism could make significant contributions to the rate of apoA-I-dependent cholesterol efflux when the expression levels of ABCA1 are low. Furthermore, the study of the effect of inhibitors of lipid efflux showed that glyburide and brefeldin A, which affect ABCA1 function, exerted strong and similar inhibitory effects on lipid efflux from both adipocytes and macrophages, whereas BLT1, an SRB-I inhibitor, only exerted a moderate inhibition. Overall these studies suggest that ABCA1 plays a major role in apoA-I-dependent lipid efflux from adipocytes and showed high similarities between the abilities of adipocytes and macrophages to release cholesterol in an apoA-I-dependent fashion.

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Abbreviations

ApoA-I:

Apolipoprotein A-I

ABCA1:

ATP-binding cassette transporter A1

GLYB:

Glyburide

BFA:

Brefeldin A

LXR:

Liver X receptor

BLT:

Block lipid transport

BSA:

Bovine serum albumin

FBS:

Fetal bovine serum

CL:

Cholesterol

PL:

Phospholipid

SR-BI:

Scavenger receptor-BI

BLT4:

Block lipid transport-4

BLT1:

Block lipid transport-1

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Acknowledgments

The authors thank Noah Barnes for his assistance in cell culture and data collection. This study was funded by Oklahoma State Experiment Station, and NIH Grant GM55622.

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  1. 147 Noble Research Center, Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK, 74078, USA

    Alisha D. Howard, Philip B. Verghese, Estela L. Arrese & Jose L. Soulages

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  1. Alisha D. Howard
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  2. Philip B. Verghese
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Correspondence to Jose L. Soulages.

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Howard, A.D., Verghese, P.B., Arrese, E.L. et al. Characterization of apoA-I-dependent lipid efflux from adipocytes and role of ABCA1. Mol Cell Biochem 343, 115–124 (2010). https://doi.org/10.1007/s11010-010-0505-7

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