本文關(guān)鍵詞：甘草總黃酮及其成分體外抗炎活性及機(jī)制研究，，由筆耕文化傳播整理發(fā)布。

甘草總黃酮及其成分體外抗炎活性及機(jī)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]上海中醫(yī)藥大學(xué)穆拉德中藥現(xiàn)代化研究中心,上海201203; [2]美國(guó)喬治華盛頓大學(xué)生物化學(xué)與分子生物學(xué)系,美國(guó)華盛頓DC20052

文章摘要：目的：考察甘草總黃酮（TFGR）及其成分對(duì)干擾素-γ（IFN-γ）和脂多糖（LPS）協(xié)同誘導(dǎo)巨噬細(xì)胞RAW26d．7炎癥模型的抗炎機(jī)制。方法：應(yīng)用溶劑萃取和大孔樹脂富集純化制備甘草醇提物、組分及總黃酮，紫外分光光度法測(cè)定總黃酮的含量；IFN—γ和LPS協(xié)同誘導(dǎo)細(xì)胞炎癥模型；Greiss反應(yīng)法測(cè)定各層次提取物及黃酮類單體對(duì)細(xì)胞上清液中亞硝酸鹽含量的影響；FRAP法測(cè)定總抗氧化能力；RT—PCR法考察TFGR及異甘草素對(duì)細(xì)胞內(nèi)誘導(dǎo)型-氧化氮合酶iNOS，COX-2，IL-1β，IL-6，PPAR-7mRNA的影響；Westernblot法考察TFGR及異甘草素對(duì)細(xì)胞iNOS，COX-2及MAPK信號(hào)轉(zhuǎn)導(dǎo)通路的影響。結(jié)果：甘草乙酸乙酯部位與其他提取物相比，在相同劑量時(shí)對(duì)亞硝酸鹽含量的抑制率最高。乙酸乙酯提取物經(jīng)大孔樹脂富集的甘草總黃酮（甘草苷占60．08％）呈劑量依賴性抑制細(xì)胞上清液中亞硝酸鹽的含量且優(yōu)于乙酸乙酯提取物，并對(duì)刺激后的細(xì)胞活力有保護(hù)作用，且總抗氧化能力也強(qiáng)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的蛋白表達(dá)；甘草黃酮類單體異甘草素能抑制iNOS，COX-2基因和蛋白表達(dá)水平和IL-1β，IL-6的基因表達(dá)，還能上調(diào)PPAR-γ的基因表達(dá)。結(jié)論：活性導(dǎo)向的分離提示乙酸乙酯部位是甘草抗炎活性組分之一，由此活性組分富集得到的TFGR呈劑量依賴性抑制細(xì)胞上清液中亞硝酸鹽含量，部分通過(guò)絲裂原活化蛋白激酶（MAPKs）中的細(xì)胞外信號(hào)調(diào)節(jié)激酶（ERK）信號(hào)通路抑制iNOS和COX-2基因和蛋白的表達(dá)而達(dá)到抗炎效果。其中異甘草素可能為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

文章關(guān)鍵詞：

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

課題項(xiàng)目：[基金項(xiàng)目]國(guó)家“重大新藥創(chuàng)制”科技重大專項(xiàng)（2009ZX09311.003）;國(guó)家自然科學(xué)基金青年基金項(xiàng)目（81001666）;上海市教委高校一氧化氮與炎癥醫(yī)學(xué)E研究院計(jì)劃項(xiàng)目（E-04010）;上海高校選拔培養(yǎng)優(yōu)秀青年教師基金項(xiàng)目（SZY0729）

  本文關(guān)鍵詞：甘草總黃酮及其成分體外抗炎活性及機(jī)制研究，由筆耕文化傳播整理發(fā)布。