红汁乳菇中愈创木烷型倍半萜化合物的抗炎活性

以新鲜红汁乳菇子实体为原料，采用有机溶剂浸提提取、乙酸乙酯萃取、硅胶柱层析得到单体化合物，通过紫外-可见光谱、高效液相色谱、质谱和核磁共振波谱分析鉴定单体化合物的结构，利用脂多糖（lipopolysaccharide，LPS）诱导RAW264.7巨噬细胞炎症模型，通过聚合酶链式反应和免疫印迹法分析单体化合物对炎症因子mRNA和蛋白相对表达水平的影响，以及对丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）炎症信号通路的作用。结果：从红汁乳菇中提取出具有较强抗炎活性的单体化合物，经鉴定为愈创木烷型倍半萜类化合物；与模型组相比，该倍半萜能极显著降低脂多糖刺激巨噬细胞中白细胞介素-6（interleukin，IL）-6、IL-1β、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和一氧化氮合酶（inducible nitric oxide synthase，iNOS）的mRNA相对表达水平（P＜0.01）；该倍半萜能降低LPS刺激的巨噬细胞中炎症因子环氧化酶-2（cyclooxygenase-2，COX-2）、IL-1β、IL-6、iNOS和TNF-α蛋白相对表达水平，且呈现一定的浓度依赖性；该倍半萜能降低磷酸化的细胞外信号激酶（p44/42）、p38 MAPK（简称p38蛋白）和c-Jun氨基端激酶（c-Jun N-terminal kinase，JNK）3 种蛋白激酶的磷酸化水平，从而抑制MAPK炎症通路的活性。

Anti-inflammatory Activity of Guaiane Sesquiterpenoids in Lactarius hatsudake

In this study, monomeric compounds from the fruiting bodies of fresh Lactarius hatsudake were obtained by sequential organic solvent extraction, ethyl acetate extraction, and silica gel column chromatography, and were structurally characterized by ultraviolet-visible (UV-Vis) spectroscopy, high performance liquid chromatography, mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. The effect of the compounds on the mRNA and protein expression of inflammatory factors and the mitogen-activated protein kinase (MAPK) signaling pathway in lipopolysaccharide (LPS)-stimulated macrophage RAW264.7 cells was determined by polymerase chain reaction (PCR) and Western blot. Results indicated that monomeric compounds with strong anti-inflammatory activity were extracted from Lactarius hatsudake and preliminarily identified as guaiane sesquiterpenoids. The guaiane sesquiterpenoids could significantly inhibit the mRNA expression of interleukin (IL)-6, IL-1β, tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) in LPS-stimulated macrophages (P < 0.01). The guaiane sesquiterpenoids could also reduce the protein expression of inflammatory factors such as cyclooxygenase-2 (COX-2), IL-1β, IL-6, iNOS and TNF-α in LPS-stimulated macrophages in a concentration-dependent manner. Therefore, the guaiane sesquiterpenoids can significantly reduce the phosphorylation levels of p44/42, p38 MAPK (p38 protein) and c-Jun N-terminal kinase (JNK), thereby inhibiting the activity of the MAPK signaling pathway.