| A sensitive assay for ABCA1-mediated cholesterol efflux using BODIPY-cholesterol. | |
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MedLine Citation:
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PMID: 21957199 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Studies have shown a negative association between cellular cholesterol efflux and coronary artery disease (CAD). Standard protocol for quantitating cholesterol efflux involves labeling cells with [3H]-cholesterol and measuring release of the labeled sterol. Using [3H]-cholesterol is not ideal for the development of a high-throughput assay to screen large numbers of serum as would be required in studying the link between efflux and CAD. We compared efflux using a fluorescent sterol (BODIPY-cholesterol) with that of [3H]-cholesterol in J774 macrophages. Fractional efflux of BODIPY-cholesterol was significantly higher than that of [3H]-cholesterol when apo A-I, HDL3, or 2% apoB-depleted human serum were used as acceptors. BODIPY-cholesterol efflux correlated significantly with [3H]-cholesterol efflux (p<0.0001) when apoB-depleted sera were used. The BODIPY-cholesterol efflux correlated significantly with preβ-1 (r2 = 0.6) but not with total HDL-C. Reproducibility of the BODIPY-cholesterol efflux assay was excellent between weeks (r2 = 0.98, inter-assay CV = 3.31%). These studies demonstrate that BODIPY-cholesterol provides an efficient measurement of efflux compared to [3H]-cholesterol and is a sensitive probe for ABCA1-mediated efflux. The increased sensitivity of BODIPY-cholesterol assay coupled with the simplicity of measuring fluorescence results in a sensitive, high-throughput assay that can screen large numbers of sera, and thus establish the relationship between cholesterol efflux and atherosclerosis. |
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Authors:
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Sandhya Sankaranarayanan; Ginny Kellner-Weibel; Margarita de la Llera Moya; Michael C Phillips; Bela F Asztalos; Robert Bittman; George H Rothblat |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-9-27 |
Journal Detail:
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Title: Journal of lipid research Volume: - ISSN: 0022-2275 ISO Abbreviation: - Publication Date: 2011 Sep |
Date Detail:
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Created Date: 2011-9-29 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0376606 Medline TA: J Lipid Res Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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Children's hospital of Philadelphia, United States; |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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