Quantifying ceramide kinetics in vivo using stable isotope tracers and LC-MS/MS.

Quantifying ceramide kinetics in vivo using stable isotope tracers and LC-MS/MS.

Chen Ying; Berejnaia Olga; Liu Jinqi; Wang Sheng-Ping; Daurio Natalie A; Yin Wu; Mayoral Rafael; Petrov Aleksandr; Kasumov Takhar; Zhang Guo-Fang; Previs Stephen F; Kelley David E; McLaren David G

American journal of physiology. Endocrinology and metabolism

2018

2018-09

Numerous studies have implicated dyslipidemia as a key factor in mediating insulin resistance. Ceramides have received special attention since their levels are inversely associated with normal insulin signaling and positively associated with factors that are involved in cardiometabolic disease. Despite the growing literature surrounding ceramide biology, there are limited data regarding the activity of ceramide synthesis and turnover in vivo. Herein, we demonstrate the ability to measure ceramide kinetics by coupling the administration of [(2)H]water with LC-MS/MS analyses. As a "proof-of-concept" we determined the effect of a diet-induced alteration on ceramide flux; studies also examined the effect of myriocin (a known inhibitor of serine palmitoyltransferase, the first step in sphingosine biosynthesis). Our data suggest that one can estimate ceramide synthesis and draw conclusions regarding the source of fatty acids; we discuss caveats in regards to method development in this area.

dyslipidemia; insulin resistance; kinetics; lipid turnover; mass spectrometry

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E416–E424

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