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 |
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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 |
format | Article |
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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.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(19)69354-3</identifier><identifier>PMID: 7228865</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>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</subject><ispartof>The Journal of biological chemistry, 1981-05, Vol.256 (10), p.4978-4983</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c410t-46407826ff66118633d129c92c35e72c709ea61c14bd4b3598c9be2a9e5363323</citedby><cites>FETCH-LOGICAL-c410t-46407826ff66118633d129c92c35e72c709ea61c14bd4b3598c9be2a9e5363323</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7228865$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Daniels, R J</creatorcontrib><creatorcontrib>Guertler, L S</creatorcontrib><creatorcontrib>Parker, T S</creatorcontrib><creatorcontrib>Steinberg, D</creatorcontrib><title>Studies on the rate of efflux of cholesterol from cultured human skin fibroblasts</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><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.</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. 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.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>7228865</pmid><doi>10.1016/S0021-9258(19)69354-3</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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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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