Quantifying apoprotein synthesis in rodents: coupling LC-MS/MS analyses with the administration of labeled water

被引:23
作者
Zhou, Haihong [1 ]
Li, Wenyu [1 ]
Wang, Sheng-Ping [1 ]
Mendoza, Vivienne [1 ]
Rosa, Raymond [1 ]
Hubert, James [1 ]
Herath, Kithsiri [1 ]
McLaughlin, Theresa [2 ]
Rohm, Rory J. [2 ]
Lassman, Michael E. [2 ]
Wong, Kenny K. [1 ]
Johns, Douglas G. [1 ]
Previs, Stephen F. [1 ]
Hubbard, Brian K. [1 ]
Roddy, Thomas P. [2 ]
机构
[1] Merck Res Labs, Atherosclerosis, Rahway, NJ 07065 USA
[2] Merck Res Labs, Vivo Pharmacoanalyt, Rahway, NJ 07065 USA
关键词
stable isotopes; liquid chromatography-tandem mass spectrometry; kinetic biomarker; dyslipidemia; cardiovascular disease; MASS ISOTOPOMER ANALYSIS; LOW-DENSITY LIPOPROTEIN; IN-VIVO MEASUREMENT; DEUTERATED WATER; PROTEIN-SYNTHESIS; AMINO-ACIDS; BODY-WATER; PLASMA-CHOLESTEROL; RAPID METHOD; SPECTROMETRY;
D O I
10.1194/jlr.D021295
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Stable isotope tracer studies of apoprotein flux in rodent models present difficulties as they require working with small volumes of plasma. We demonstrate the ability to measure apoprotein flux by administering either H-2- or O-18-labeled water to mice and then subjecting samples to LC-MS/MS analyses; we were able to simultaneously determine the labeling of several proteolytic peptides representing multiple apoproteins. Consistent with relative differences reported in the literature regarding apoprotein flux in humans, we found that the fractional synthetic rate of apoB is greater than apoA1 in mice. In addition, the method is suitable for quantifying acute changes in protein flux: we observed a stimulation of apoB production in mice following an intravenous injection of Intralipid and a decrease in apoB production in mice treated with an inhibitor of microsomal triglyceride transfer protein. In summary, we demonstrate a high-throughput method for studying apoprotein kinetics in rodent models. Although notable differences exist between lipoprotein profiles that are observed in rodents and humans, we expect that the method reported here has merit in studies of dyslipidemia as i) rodent models can be used to probe target engagement in cases where one aims to modulate apoprotein production and ii) the approach should be adaptable to studies in humans.-Zhou, H., W. Li, S-P. Wang, V Mendoza, R. Rosa, J. Hubert, K. Herath, T. McLaughlin, R. J. Rohm, M. E. Lassman, K. K. Wong, D. G. Johns, S. F. Previs, B. K. Hubbard, and T. P. Roddy. Quantifying apoprotein synthesis in rodents: coupling LC-MS/MS analyses with the administration of labeled water. J. Lipid Res. 2012. 53: 1223-1231.
引用
收藏
页码:1223 / 1231
页数:9
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