| RRC ID |
59994
|
| Author |
Shimizu T, Miyazaki O, Iwamoto T, Usui T, Sato R, Hiraishi C, Yoshida H.
|
| Title |
A new method for measuring cholesterol efflux capacity uses stable isotope-labeled, not radioactive-labeled, cholesterol.
|
| Journal |
J Lipid Res
|
| Abstract |
The incidence of cardiovascular events correlates inversely with cholesterol efflux capacity (CEC) more than with HDL-cholesterol level. The measurement of CEC is used to qualify cardiovascular disease risk and is conventionally performed with radioisotope (RI)-labeled cholesterol. Here, we established a CEC measurement technique using stable isotope-labeled cholesterol as an alternative, and we compared the new method with RI and fluorescence (boron dipyrromethene difluoride-cholesterol) methods in cells and in patient serum. We incubated J774 cells labeled with [d7]cholesterol ([d7]C) with patient serum depleted of apoB, and [d7]C extracted from the culture medium was quantified by liquid chromatography/quadrupole time-of-flight mass spectrometry. [d7]C efflux increased with greater apoB-depleted serum concentration and longer incubation time. The assay coefficient of variation (CV) of five consecutive measurements of three sets of samples ranged from 7.3% to 9.5%, and the interassay CV determined by measuring three samples four times ranged from 4.1% to 8.5%, both indicating good precision. We then measured CEC levels of 41 outpatients with serum HDL-cholesterol levels between 36 and 94 mg/dl (mean: 61.7 ± 18.0 mg/dl); in the presence of cAMP, we observed a significant, positive correlation between CEC levels determined with the stable isotope and RI methods that was stronger than the correlation between measurements obtained by the fluorescence and RI methods (r = 0.73, P < 0.0001 vs. r = 0.55, P < 0.001). Therefore, our stable isotope method can be considered useful as a non-RI method and thus deserves evaluation in future clinical studies.
|
| Volume |
60(11)
|
| Pages |
1959-1967
|
| Published |
2019-11-1
|
| DOI |
10.1194/jlr.D086884
|
| PII |
S0022-2275(20)32294-X
|
| PMID |
31455616
|
| PMC |
PMC6824490
|
| MeSH |
Adult
Aged
Aged, 80 and over
Animals
Capillary Electrochromatography / methods*
Cell Line
Cholesterol, HDL / blood*
Female
Humans
Isotope Labeling / methods
Male
Mice
Middle Aged
|
| IF |
4.743
|
| Times Cited |
0
|
| Resource |
| Human and Animal Cells |
J774.1(RCB0434) |
