Studies on the rate of efflux of cholesterol from cultured human skin fibroblasts

The cholesterol content of normal human skin fibroblasts was increased (approximately doubled) by incubating cells in the presence of a high concentration of low density lipoprotein. Cholesterol efflux from these cells was then studied as a function of time and as a function of acceptor concentratio...

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Veröffentlicht in:The Journal of biological chemistry 1981-05, Vol.256 (10), p.4978-4983
Hauptverfasser: Daniels, R J, Guertler, L S, Parker, T S, Steinberg, D
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container_end_page 4983
container_issue 10
container_start_page 4978
container_title The Journal of biological chemistry
container_volume 256
creator Daniels, R J
Guertler, L S
Parker, T S
Steinberg, D
description The cholesterol content of normal human skin fibroblasts was increased (approximately doubled) by incubating cells in the presence of a high concentration of low density lipoprotein. Cholesterol efflux from these cells was then studied as a function of time and as a function of acceptor concentration. High density lipoprotein from which essentially all of the cholesterol had been removed by heptane extraction was used as a model acceptor (cholesterol-depleted high density lipoprotein). Using a sensitive enzymatic assay, it was possible to measure the increase in medium cholesterol and the decrease in cell cholesterol content simultaneously. Release was approximately a linear function of time for at least 6-12 h. A maximal rate of release was obtained at 20 micrograms of protein/ml (50% of excess stored sterol released in about 12 h); increasing the acceptor concentration 10-fold (to 200 micrograms/ml) failed to increase efflux rate. Comparison of the rates of fall of free and ester cholesterol levels suggested that hydrolysis of the ester may be rate-limiting when cholesterol-depleted high density lipoprotein is used as the acceptor. The results imply that above saturating concentrations of acceptor, acceptor-cell interaction is no longer limiting and that the rate of efflux of cholesterol under such conditions depends on intracellular processes necessary to make cholesterol available to the acceptor (e.g. hydrolysis of cholesterol esters and transfer of cholesterol from intracellular sites to the plasma membrane). Whether or not the concentrations of acceptor bathing cells in vivo is normally rate-limiting remains to be determined.
doi_str_mv 10.1016/S0021-9258(19)69354-3
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The results imply that above saturating concentrations of acceptor, acceptor-cell interaction is no longer limiting and that the rate of efflux of cholesterol under such conditions depends on intracellular processes necessary to make cholesterol available to the acceptor (e.g. hydrolysis of cholesterol esters and transfer of cholesterol from intracellular sites to the plasma membrane). 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Cholesterol efflux from these cells was then studied as a function of time and as a function of acceptor concentration. High density lipoprotein from which essentially all of the cholesterol had been removed by heptane extraction was used as a model acceptor (cholesterol-depleted high density lipoprotein). Using a sensitive enzymatic assay, it was possible to measure the increase in medium cholesterol and the decrease in cell cholesterol content simultaneously. Release was approximately a linear function of time for at least 6-12 h. A maximal rate of release was obtained at 20 micrograms of protein/ml (50% of excess stored sterol released in about 12 h); increasing the acceptor concentration 10-fold (to 200 micrograms/ml) failed to increase efflux rate. Comparison of the rates of fall of free and ester cholesterol levels suggested that hydrolysis of the ester may be rate-limiting when cholesterol-depleted high density lipoprotein is used as the acceptor. The results imply that above saturating concentrations of acceptor, acceptor-cell interaction is no longer limiting and that the rate of efflux of cholesterol under such conditions depends on intracellular processes necessary to make cholesterol available to the acceptor (e.g. hydrolysis of cholesterol esters and transfer of cholesterol from intracellular sites to the plasma membrane). Whether or not the concentrations of acceptor bathing cells in vivo is normally rate-limiting remains to be determined.</description><subject>Blood</subject><subject>cell culture</subject><subject>Cells, Cultured</subject><subject>cholesterol</subject><subject>Cholesterol - metabolism</subject><subject>Culture Media</subject><subject>fibroblasts</subject><subject>Fibroblasts - metabolism</subject><subject>Humans</subject><subject>Infant</subject><subject>Kinetics</subject><subject>Lipoproteins, HDL - metabolism</subject><subject>Lipoproteins, LDL - metabolism</subject><subject>Male</subject><subject>man</subject><subject>skin</subject><subject>Skin - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1981</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtLxDAQgIMouj5-ghAQRA_VTF5tjiK-QBBRwVto04mtthtNWtR_b9ddvDqXGZhvHnyE7AM7AQb69IExDpnhqjgCc6yNUDITa2QGrBCZUPC8TmZ_yBbZTumVTSENbJLNnPOi0GpG7h-GsW4x0TCnQ4M0lgPS4Cl6341fi8o1ocM0YAwd9TH01I3dMEasaTP25Zymt3ZOfVvFUHVlGtIu2fBll3BvlXfI0-XF4_l1dnt3dXN-dps5CWzIpJYsL7j2XmuAQgtRAzfOcCcU5tzlzGCpwYGsalkJZQpnKuSlQSUmmIsdcrjc-x7Dxzh9aPs2Oey6co5hTDZXWnI9yfgPBDU5YVpNoFqCLoaUInr7Htu-jN8WmF04t7_O7UKoBWN_nVsxze2vDoxVj_Xf1Ery1D9Y9pv2pflsI9qqDa7B3nKlF7ulyQvxA-oyh14</recordid><startdate>19810525</startdate><enddate>19810525</enddate><creator>Daniels, R J</creator><creator>Guertler, L S</creator><creator>Parker, T S</creator><creator>Steinberg, D</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FD</scope><scope>FR3</scope><scope>M7Z</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19810525</creationdate><title>Studies on the rate of efflux of cholesterol from cultured human skin fibroblasts</title><author>Daniels, R J ; Guertler, L S ; Parker, T S ; Steinberg, D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c410t-46407826ff66118633d129c92c35e72c709ea61c14bd4b3598c9be2a9e5363323</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1981</creationdate><topic>Blood</topic><topic>cell culture</topic><topic>Cells, Cultured</topic><topic>cholesterol</topic><topic>Cholesterol - metabolism</topic><topic>Culture Media</topic><topic>fibroblasts</topic><topic>Fibroblasts - metabolism</topic><topic>Humans</topic><topic>Infant</topic><topic>Kinetics</topic><topic>Lipoproteins, HDL - metabolism</topic><topic>Lipoproteins, LDL - metabolism</topic><topic>Male</topic><topic>man</topic><topic>skin</topic><topic>Skin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Daniels, R J</creatorcontrib><creatorcontrib>Guertler, L S</creatorcontrib><creatorcontrib>Parker, T S</creatorcontrib><creatorcontrib>Steinberg, D</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Daniels, R J</au><au>Guertler, L S</au><au>Parker, T S</au><au>Steinberg, D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Studies on the rate of efflux of cholesterol from cultured human skin fibroblasts</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1981-05-25</date><risdate>1981</risdate><volume>256</volume><issue>10</issue><spage>4978</spage><epage>4983</epage><pages>4978-4983</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The cholesterol content of normal human skin fibroblasts was increased (approximately doubled) by incubating cells in the presence of a high concentration of low density lipoprotein. 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ispartof The Journal of biological chemistry, 1981-05, Vol.256 (10), p.4978-4983
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source MEDLINE; Alma/SFX Local Collection; EZB Electronic Journals Library
subjects Blood
cell culture
Cells, Cultured
cholesterol
Cholesterol - metabolism
Culture Media
fibroblasts
Fibroblasts - metabolism
Humans
Infant
Kinetics
Lipoproteins, HDL - metabolism
Lipoproteins, LDL - metabolism
Male
man
skin
Skin - metabolism
title Studies on the rate of efflux of cholesterol from cultured human skin fibroblasts
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