Non-targeted Metabolomics Intergrated with 1 H NMR and LC-Q-TOF-MS/MS Methods to Depict a More Comprehensive Metabolic Profile in Response to Chrysosplenetin and Artemisinin Co-Treatment against Artemisinin-Sensitive and -Resistant Plasmodium berghei K173.

Our previous work revealed mutual and specific metabolites/pathways in artemisinin sensitive and resistant Plasmodium berghei K173 infected mice. In this study, we further investigated whether chrysosplenetin, a candidate chemical to prevent artemisinin resistance, can regulate these metabolites/pathways by integrating non-targeted metabolomics with 1 H NMR and LC-Q-TOF-MS/MS spectrum. NMR method generated specifically altered metabolites in response to co-treatment with chrysosplenetin, including the products of glycolysis such as glucose, pyruvate, lactate, and alanine; taurine, closely associated with liver injury; arginine and proline as essential amino acids for parasites; TMAO, a biomarker for dysbacteriosis and renal function; and tyrosine, which is used to generate levodopa and dopamine and may improve the torpor state of mice. Importantly, we noticed that chrysosplenetin might depress the activated glycolysis induced by sensitive parasites, but oppositely promoted the inhibited glycolysis to generate more lactate which suppresses proliferation of resistant parasites. Moreover, chrysosplentin possibly disturb heme biosynthetic pathway in mitochondria. MS method yielded changed coenzyme A (CoA), phosphatidylcholine (PC), and ceramides (cer), closely related to mitochondria β-oxidation, cell proliferation, differentiation, and apoptosis. These two means shared no overlapped metabolites and formed a more broader metabolic map to study the potential mechanisms of chrysosplenetin as a promising artemisinin resistance inhibitor.