Monitoring carnosine uptake by RAW 264.7 macrophage cells using microchip electrophoresis with fluorescence detection
journal contributionposted on 07.03.2019 by Claudia G. Fresta, Michael L. Hogard, Giuseppe Caruso, Elton E. Melo Costa, Giuseppe Lazzarino, Susan M. Lunte
Any type of content formally published in an academic journal, usually following a peer-review process.
In this report, a microchip electrophoresis system with fluorescence detection was used for the quantitation of intracellular carnosine in untreated and stimulated macrophage cell lysates. Carnosine was derivatized with NDA/CN and separated from other endogenous amine reported in macrophage cells. Based on ME-LIF with standard addition, macrophages were estimated to contain a basal intracellular concentration of carnosine (0.079 ± 0.02 nmol per million cells). Carnosine is readily taken up by macrophages in cell culture. Incubation with 20 mM carnosine led to a 600-fold increase in intracellular carnosine compared to basal levels. Furthermore, we have shown that under pro-inflammatory conditions using LPS and IFN-gamma stimulation there is a further 3-fold increase in carnosine uptake in macrophage cells. This suggests that there is a mechanism through which macrophages increase the usage of carnosine during oxidative stress.