total flavonoids of Glycyrrhizae Radix et Rhizoma macrophage

  本文关键词：甘草总黄酮及其成分体外抗炎活性及机制研究，，由笔耕文化传播整理发布。

甘草总黄酮及其成分体外抗炎活性及机制研究

Study on in vitro anti-inflammatory activity of total flavonoids from Glycyrrhizae Radix et Rhizoma and its ingredients

[1] [2] [3] [4]

YANG Xiao-lu , LIU Duo , BIAN Ka, ZHANG Dan-dan （ 1. Murad Research Institute for Modernized Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai

[1]上海中医药大学穆拉德中药现代化研究中心,上海201203; [2]美国乔治华盛顿大学生物化学与分子生物学系,美国华盛顿DC20052

文章摘要：目的：考察甘草总黄酮（TFGR）及其成分对干扰素-γ（IFN-γ）和脂多糖（LPS）协同诱导巨噬细胞RAW26d．7炎症模型的抗炎机制。方法：应用溶剂萃取和大孔树脂富集纯化制备甘草醇提物、组分及总黄酮，紫外分光光度法测定总黄酮的含量；IFN—γ和LPS协同诱导细胞炎症模型；Greiss反应法测定各层次提取物及黄酮类单体对细胞上清液中亚硝酸盐含量的影响；FRAP法测定总抗氧化能力；RT—PCR法考察TFGR及异甘草素对细胞内诱导型-氧化氮合酶iNOS，COX-2，IL-1β，IL-6，PPAR-7mRNA的影响；Westernblot法考察TFGR及异甘草素对细胞iNOS，COX-2及MAPK信号转导通路的影响。结果：甘草乙酸乙酯部位与其他提取物相比，在相同剂量时对亚硝酸盐含量的抑制率最高。乙酸乙酯提取物经大孔树脂富集的甘草总黄酮（甘草苷占60．08％）呈剂量依赖性抑制细胞上清液中亚硝酸盐的含量且优于乙酸乙酯提取物，并对刺激后的细胞活力有保护作用，且总抗氧化能力也强于其他提取物；TFGR抑制iNOS，IL-6mRNA并提高PPAR—γ基因水平，抑制iNOS，COX-2，β-ERK的蛋白表达；甘草黄酮类单体异甘草素能抑制iNOS，COX-2基因和蛋白表达水平和IL-1β，IL-6的基因表达，还能上调PPAR-γ的基因表达。结论：活性导向的分离提示乙酸乙酯部位是甘草抗炎活性组分之一，由此活性组分富集得到的TFGR呈剂量依赖性抑制细胞上清液中亚硝酸盐含量，部分通过丝裂原活化蛋白激酶（MAPKs）中的细胞外信号调节激酶（ERK）信号通路抑制iNOS和COX-2基因和蛋白的表达而达到抗炎效果。其中异甘草素可能为TFGR抗炎的活性成分。

Abstr：Objective： To examine the anti-inflammatory mechanism of total flavonoids of Glycyrrhizae Radix et Rhizoma（ TF- GR） and its ingredient on IFN-T and LPS-induced macrophage RAW264. 7. Method： Solvent extraction and macroporous resin enrich- ment were adopted for preparing ethanol extracts of Glycyrrhizae Radix et Rhizoma, components and total flavonoids. Ultraviolet spec- troscopy was used to determine the content of total flavonoids. An IFN-T and LPS-induced cell inflammatory model was established. Griess reaction was used for detecting the effect of extracts at all levels and ftavonoid monomers on nitrite content in cell culture super- natant. FRAP was used for measuring anti-oxidation capacity. RT-PCR was used for determining the effect of TFGR and isoliquiriti- genins on intracellular inducible nitric oxide synthase iNOS, COX-2, IL-6 and PPAR-γ. Western blot was used for detecting the effect of TFGR and isoliquiritigenins on iNOS, COX-2 and MAPK signal transduction pathways. Result： Compared with other extracts, ethyl acetate fractions from Glyeyrrhizae Radix et Rhizoma showed the highest inhibition ratio on nitrite content at the same concentration. After being enriched with maeroporous resin, TFGR （60. 08% of liquiritin） of ethyl acetate extracts from Glycyrrhizae Radix et Rhizo- ma showed dose-dependence, and inhibited the nitrite content in cell culture supernatant, which was superior to ethyl acetate extracts, and had the protective effect on post-stimulated cell activity, with a stronger total anti-oxidation than other extracts. TFGR inhibited iN- OS, IL-6 mRNA, protein expressions of iNOS, COX-2 and IL-6. Isoliquiritigenin, a flavonoid monomer, could inhibited iNOS, COX- 2 gene and protein expression and gene

文章关键词：

Keyword：:total flavonoids of Glycyrrhizae Radix et Rhizoma macrophage inflammation mechanism of action

课题项目：[基金项目]国家“重大新药创制”科技重大专项（2009ZX09311.003）;国家自然科学基金青年基金项目（81001666）;上海市教委高校一氧化氮与炎症医学E研究院计划项目（E-04010）;上海高校选拔培养优秀青年教师基金项目（SZY0729）

  本文关键词：甘草总黄酮及其成分体外抗炎活性及机制研究，由笔耕文化传播整理发布。