Metabolic profiling of culture medium of brain microvascular endothelial cells challanged with neospora caninum
This study aimed to assess if the pathophysiological impact of N. caninum infection on cultured endothelial cells could be explained by changes in the nutritional needs of host cells during the course of intracellular (i.c.) infection. Fluctuations of culture medium nutrients in response to growth of N. caninum were determined using three approaches. First, concentrations of several extracellular biochemical parameters of Brain Microvascular Endothelial Cell (BMEC) culture medium were monitored in a time course as a function of infection using a Randox RX Imola clinical chemistry analyser. Second, extracellular metabolite changes in culture medium were monitored by using Raman spectroscopy (RS). Third, a targeted LC-MS method was employed to quantify important metabolite groups like amino acids, organic acids, sugar phosphates, other phosphorylated metabolites, and nucleotides in the medium of infected cell cultures and control cultures at 24 hr pi. Cell viability was determined by LDH assay and demonstrated that N. caninum caused cytotoxicity in endothelial cells in a dose-response manner. Beta hydroxybutyrate, pyruvate, ATP, total protein (tp), non-esterified fatty acids (NEFA), and triglycerides were significantly different in infected cells compared with controls. RS and metabolite profiling using LC-MS/MS revealed alteration in certain metabolites.
