桑白皮中sanggenon B对脂多糖诱导RAW264.7细胞炎症反应的影响  

Effect of Sanggenon B from Cortex Mori on Lipopolysaccharide-induced Inflammatory Response in RAW264.7 Cells

作　　者：吴永祥[1] 吴丽萍[1] 胡长玉[1] 万志兵[1] 李丰伯[1] 金泰完 Yong-xiang;WU Li-ping;HU Chang-yu;WAN Zhi-bing;LI Feng-bo;KIM Tae-wan(College of Limb and Environment Science,Huangshan University,Huangshan 245041,China;Department of Food Scienee and Biotechnology,Andong National University,Andong,760749,Korea)

机构地区：[1]黄山学院生命与环境科学学院,黄山245041 [2]韩国安东国立大学食品科学与生物技术学院,安东760749

出　　处：《天然产物研究与开发》2018年第7期1132-1137,共6页Natural Product Research and Development

基　　金：安徽省高校自然科学研究重点项目(KJ2017A398);安徽省留学回国人员创新项目择优资助计划重点项目(2017srst1)

摘　　要：采用硅胶柱层析等方法从桑白皮中分离出抗炎活性化合物,通过波谱分析进行化合物结构鉴定。体外培养RAW264.7细胞,构建脂多糖(LPS)诱导炎症模型,采用噻唑蓝(MTT)法测定细胞活性,利用Griess法和酶联免疫吸附(ELISA)法分别检测炎症因子一氧化氮(NO)和肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)的释放量,采用实时荧光定量PCR(qRT-PCR)和Western blot技术分别测定诱导型一氧化氮合酶(iNOS)、环氧合酶-2(COX-2)mRNA和蛋白的表达水平。结果显示:桑白皮乙醇提取物(MRE)能显著抑制LPS诱导的RAW264.7细胞NO的分泌水平,且呈现量效关系。对桑白皮进行分离,得到1种抗炎活性化合物,经波谱数据分析,确定其为已知的Diels-Alder加合物桑根酮B(sanggenon B)。抗炎活性表明,sanggenon B显著下调了炎症因子NO、TNF-α、IL-6的合成,并抑制了RAW264.7细胞中i NOS、COX-2 mRNA和蛋白的表达水平。研究表明,从桑白皮中分离出的sanggenon B具有一定的抗炎作用,其抗炎机制可能与调控炎症因子的合成,降低i NOS、COX-2表达有关。Cortex Mori was separated by silica gel column chromatography to afford the active compound. The structure elucidation of the compound was identified by analysis of spectroscopic data. RAW264. 7 cells were stimulated by lipopolysaccharide（ LPS） to induce inflammatory model. Cell viability was determined using a methyl thiazolyl tetrazolium（ MTT） assay. The production of nitric oxide（ NO） and tumor necrosis factor-α（ TNF-α）,interleukin 6（ IL-6） were measured by Griess reaction and ELISA,respectively. The inducible nitric oxide synthase（ i NOS） and cyclooxygenase-2（ COX-2） mRNA and protein levels were measured by qRT-PCR and Western blot,respectively. Results showed Cortex Mori ethanol extract（ MRE） was able to inhibit the production of NO in LPS-stimulated RAW264. 7 cells,in a dose-dependent manner. One active compound was isolated from Cortex Mori and identified as the Diels-Alder type adducts sanggenon B. The anti-inflammatory results showed that sanggenon B could significantly inhibit the production of NO,TNF-α,and IL-6. Sanggenon B also caused a significant dose-dependent decrease in the mRNA and protein expressions of iNOS,COX-2. In conclusion,sanggenon B isolated from Cortex Mori exhibited obvious anti-inflammatory effect and its mechanism may be related to its regulation on inflammatory cytokines,and inhibition of the expression of i NOS,COX-2.

关 键 词：桑白皮 桑根酮B RAW264.7细胞 炎症 Diels-Alder加合物 

分 类 号：R284.1[医药卫生—中药学]