Novel in vivo method for measuring cholesterol mass flux in peripheral macrophages.
Novel in vivo method for measuring cholesterol mass flux in peripheral macrophages.
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DOI：
10.1161/atvbaha.111.236406
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发表时间：
2011-12
期刊：
影响因子：
--
通讯作者：
Rothblat GH
中科院分区：
文献类型：
--
作者：
Weibel GL;Hayes S;Wilson A;Phillips MC;Billheimer J;Rader DJ;Rothblat GH
Reverse cholesterol transport (RCT) is the process by which excess cholesterol is removed from peripheral tissue by high density lipoprotein (HDL) and delivered to the liver for excretion. Presently, methods of measuring in vivo RCT do so by monitoring the appearance in the feces of labeled cholesterol that originated from peripheral macrophage foam cells. These methods do not account for changes in macrophage cholesterol mass. We have developed an in vivo assay to measure cholesterol mass changes in atherosclerotic foam cells. Macrophages are entrapped in semi-permeable (pore size 0.2μm) hollow fibers and surgically implanted into the peritoneum of recipient mice. The fibers are removed from the peritoneum 24h after implantation. This method allows the complete recovery of the macrophages for quantification of changes in cholesterol mass and cellular protein. In wild type mice we measured a significant reduction in total cell cholesterol (TC) when hollow fibers containing cholesterol-enriched macrophage cells were implanted (TC before implantation = 105±18μg/mg cell protein, TC 24h after implantation = 60±16μg/mg protein). Additionally, there was an increase in cholesterol content when hollow fibers containing cholesterol-normal macrophages were implanted in an atherogenic mouse model (LDLr/apobec dko) compared to a wild type mouse (initial TC content = 57±24μg/mg protein, TC 24h after implantation: wild type mice = 52±10μg/mg protein; LDLr/apobec dko mice = 118±27μg/mg protein). This assay can quantify in vivo both cholesterol mass accumulation, and reduction, in macrophages. This method permits quantitative analysis of the progression and regression of foam cells.
