Abstract:In this study, a ceramides-dominant mixture was isolated from Incarvillea compacta by silica gel column chromatography and identified by ultra performance liquid chromatography tandem mass spectrometry along with identification methods such as H/C nuclear magnetic resonance spectroscopy. The ceramides-dominant mixture had a total ceramide content of 81.53%, and its anti-inflammatory effect was preliminarily evaluated. Besides, the anti-inflammatory effect of the total ceramides from I. compacta on (LPS)-stimulated RAW264.7 cells were also investigated. The results of methyl thiazolyl tetrazolium (MTT) assay showed that the ceramides-dominant mixture from I. compacta (the ceramide preparation) at low concentrations (6.25~50 μg/mL) had no significant effect on the cell viability of RAW264.7 cells. The ceramide preparation significantly inhibited the lipopolysaccharide (LPS)-induced production of nitric oxide (NO), with the highest inhibition rate being 91.07%±0.11% when the concentration of the ceramide preparation was 100 μg/mL. The ceramide preparation effectively suppressed the mRNA expressions of interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOs), as well as the protein expressions and phosphorylation of inhibitor-α of NF-κB (IκBα), pyruvate dehydrogenase kinase1 (PDK1), protein kinase B (AKT) in LPS-stimulated RAW264.7 cells. The results of this study showed that the ceramide preparation has significant anti-inflammatory activity, and can exert its anti-inflammatory effect through phosphatidylinositol 3-kinase / protein kinase B (PI3K/AKT) signaling pathway. The experimental results of this work can provide certain scientific and data support for the development of related health foods and rational utilization of plant resources.
