虾青素DHA单酯对脂多糖诱导炎症反应的保护作用

Protective Effect of Astaxanthin Docosahexaenoic Acid Monoester on Lipopolysaccharide-Induced Inflammatory Responses

目的:通过构建脂多糖(lipopolysaccharide,LPS)诱导的BV2小鼠小胶质细胞和斑马鱼炎症模型研究虾青素二十二碳六烯酸(docosahexaenoic,DHA)单酯的抗炎活性。方法:采用光学显微镜观察细胞形态,四甲基偶氮唑蓝法检测细胞活性,Griess法测定NO水平。向斑马鱼胚胎显微注射虾青素,体视显微镜下观察胚胎发育,采用试剂盒检测胚胎中活性氧(reactive oxygen species,ROS)产生量和丙二醛(malondialdehyde,MDA)含量;采用实时荧光定量聚合酶链式反应测定炎症因子的表达水平。结果:虾青素DHA单酯干预后,细胞内炎症因子表达水平与模型组相比极显著下降(P<0.01),接近于空白对照组;游离虾青素和虾青素DHA单酯均能保护斑马鱼胚胎的发育,降低LPS诱导的胚胎畸形率,有效抑制炎症斑马鱼体内ROS产生,同时显著降低MDA含量,下调炎症因子的表达水平,并且虾青素DHA单酯的效果更佳。结论:虾青素DHA单酯比游离虾青素具有更好的抗炎活性,对LPS诱导的炎症反应有明显的保护作用。

Purpose: The anti-inflammatory activity of astaxanthin docosahexaenoic acid(DHA) monoester was investigated in lipopolysaccharide(LPS)-induced mouse microglial BV2 cells and zebrafish. Methods: Cell morphology was observed by light microscope, cell activity was detected by methyl thiazolyl tetrazolium assay, and nitric oxide(NO) level was determined by Griess method. Zebrafish embryos were microinjected with astaxanthin, and embryo development was observed under a stereomicroscope. The contents of reactive oxygen species(ROS) and malondialdehyde(MDA) were detected with commercial kits. The expression levels of inflammatory factors were determined by real-time fluorescence quantitative polymerase chain reaction. Results: After astaxanthin DHA monomer intervention, the expression level of intracellular inflammatory factors decreased significantly compared with the model group, approaching that in the blank control group. Both free astaxanthin and astaxanthin DHA monomer could protect the development of zebrafish embryos,lower the incidence of embryo malformations induced by LPS, effectively inhibit ROS production in zebrafish with inflammation, significantly reduce the content of MDA, and down-regulate the expression levels of inflammatory factors,and the effect of astaxanthin DHA monomer was more pronounced. Conclusion: Astaxanthin DHA monoester has a stronger anti-inflammatory activity, and a more significant protective effect against LPS-induced inflammation than free astaxanthin.