密生波罗花中神经酰胺类成分的抗炎作用

Anti-inflammatory Effects of Ceramides from Incarvillea compacta

该研究运用硅胶柱层析法,结合超高效液相色谱-高分辨串联质谱联用技术以及核磁共振氢谱/碳谱等鉴定方法,从密生波罗花中分离鉴定了一组以神经酰胺为主的混合物,其中,神经酰胺类成分含量为81.53%,并对其抗炎作用进行了初步评价。噻唑蓝(MTT)检测结果显示低浓度的密生波罗花神经酰胺(6.25~50μg/mL)对RAW264.7的细胞活性没有显著影响。密生波罗花神经酰胺能显著减少脂多糖(LPS)诱导RAW264.7细胞一氧化氮(NO)的释放量,且当浓度为100μg/mL时,对NO释放的抑制率最高,为91.07%±0.11%。密生波罗花神经酰胺能高效抑制LPS诱导RAW264.7细胞中白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)及一氧化氮合酶(iNOS)等炎症因子的mRNA表达水平,显著降低LPS诱导RAW264.7细胞中核因子κB抑制蛋白α(IκBα)、丙酮酸脱氢酶激酶1(PDK1)及AKT蛋白的表达及磷酸化水平。结果表明密生波罗花神经酰胺具有显著的抗炎活性,并可通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路发挥其抗炎作用,可为相关保健食品开发和该植物资源的合理利用提供一定的科学依据和数据支撑。

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(6.25~50μg/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 100μg/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-αof 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.