赤芝FPS基因酵母单杂交文库构建及其上游转录因子的筛选

目的筛选赤芝三萜合成途径中法尼基焦磷酸合酶(farnesyl diphosphate synthase,FPS)基因的上游调控转录因子。方法首先克隆了FPS基因启动子,并连接至pAbAi质粒构建诱饵载体pAbAi-FPS。将pAbAi-FPS转化Y1H酵母感受态细胞构建诱饵菌。采用SMART技术构建赤芝酵母单杂交cDNA文库,再将纯化的双链cDNA、pGADT7-Rec共转化诱饵菌株,筛选FPS上游的转录调控因子。结果构建了含pAbAi-FPS的诱饵载体并筛选出诱饵菌株,构建了cDNA文库并转化诱饵菌株,筛选出阳性克隆37个,得到作用FPS基因上游的转录调控因子18个,包括转录因子3个、核糖体蛋白5个及其他家族蛋白10个。结论筛选出转录因子GlSNF2、GlMHR和GlZn2Cys6为调控FPS表达的候选基因,为深入研究FPS基因的表达调控机制奠定了研究基础。     

藏族药西伯利亚蓼化学成分的分离鉴定

目的:对蓼科Polygonaceae植物藏族药西伯利亚蓼Polygonum sibiricum的干燥全草进行化学成分研究。方法:藏族药西伯利亚蓼的干燥全草用70%乙醇提取,浓缩,过大孔树脂,依次用水,30%乙醇,50%乙醇,90%乙醇洗脱,对90%乙醇洗脱部分采用各种柱色谱(硅胶,LH-20羟丙基葡聚糖凝胶,反相色谱Rp-18)进行分离,通过波谱数据分析(1H-NMR,13CNMR,MS)进行结构鉴定。结果:从90%乙醇部分分离鉴定了8个化合物,其中4个黄酮类化合物,1个酚性化合物,1个三萜化合物以及2个甾体化合物,分别鉴定为槲皮素(quercetin,1),山柰酚(kaempferol,2),木犀草素(luteolin,3),番石榴苷(guajavarin,4),邻羟基苯甲酸(o-hydroxybenzoic acid,5),齐墩果酸(oleanolic acid,6),胡萝卜苷(daucosterol,7),谷甾醇(β-sitosterol,8)。结论:化合物1~8均为首次从藏族药西伯利亚蓼中分离得到,该研究为寻找藏族药西伯利亚蓼抗痛风药效物质提供一定的化学依据。     

小花清风藤化学成分的研究

目的研究小花清风藤Sabia parviflora茎叶的化学成分。方法利用硅胶、Sephadex LH-20、反相硅胶、MCI等柱色谱以及半制备高效液相色谱等多种色谱技术进行分离纯化,运用NMR、MS等波谱学方法以及结合文献数据鉴定化合物结构。结果从小花清风藤茎叶的石油醚萃取部位分离得到14个化合物,包括7个五环三萜类化合物,4个生物碱类化合物,3个其他类化合物;分别鉴定为β-谷甾醇(1)、木栓酮(2)、(20S)-3-oxo-20-hydroxytaraxastane(3)、9-芴酮(4)、N-formyldehydroanonain(5)、桦木酸(6)、dehydroformouregine(7)、棕榈酸(8)、20-hydroxy-lupan-3-one(9)、3-氧代齐墩果酸(10)、古柯三萜二醇(11)、桦木酸甲酯(12)、N-formyl-annonain(Z)(13)、N-formyl-O-methylisopiline(14)。结论化合物3~10、12~14为首次从该植物以及该属植物中得到,首次报道了化合物5、7、14的~(13)C-NMR谱归属。     

牛白藤的化学成分及其免疫抑制活性研究

研究茜草科耳草属植物牛白藤Hedyotis hedyotidea(DC.) Merr.干燥藤茎的化学成分及免疫抑制活性,为进一步开发利用牛白藤提供依据.以硅胶、凝胶柱色谱、制备高效液相色谱进行分离纯化,并应用MS和NMR技术分析鉴定化合物结构,分离得到11个化合物吐叶醇(6S,9S-vomifoliol,1),桦木酮酸(betulonic acid,2),白桦脂酸(betulinic acid,3),白桦醇(betulin,4),表白桦脂酸(3-epi-betulinic acid,5),乌苏酸(ursolic acid,6),β-谷甾醇(β-sitosterol,7),stigmast-4-en-3-one (8),7β-hydroxysitosterol(9),(3β,7β)-7-methoxystigmast-5-en-3-ol(10)和巴戟醚萜(morindacin,11).化合物1,2,4,8～ 11均首次从该植物中分得,化合物1,2,8 ～ 11均首次从耳草属植物中分得,化合物9,10均首次从茜草科植物中分得.通过体外淋巴细胞转化实验测试化学成分的免疫抑制活性,化合物4,6,9均有较强的免疫抑制活性.     

Triterpenes from Astilbe chinensis

Six new triterpenes, 3β,6β-dihydroxyurs-12-en-27-oic acid (1), 3β,6β,24-trihydroxyurs-12-en-27-oic acid (2), 3β,6β,7α-trihydroxyurs-12-en-27-oic acid (3), 3β-acetoxy-6β-hydroxyurs-12-en-27-oic acid (4), 3β,6β,24-trihydroxyolean-12-en-27-oic acid (5), and 3β,6β,7α-trihydroxyolean-12-en-27-oic acid (6), were isolated from the rhizomes of Astilbe chinensis. Their structures were elucidated on the basis of 1D- and 2D-NMR analyses and HR-MS experiments. The isolated compounds exhibited significant cytotoxic activities against the SK-N-SH and HL-60 cell lines.     

维吾尔族药大戟脂中三萜类化学成分研究

该研究通过反复硅胶柱色谱、ODS柱色谱和半制备型HPLC等方法,从维吾尔族药大戟脂的甲醇超声提取物中共分离得到10个三萜类化合物。利用各种波谱学手段及理化性质确定了这些化合物的结构,分别鉴定为3β-hydroxy-25,26,27-trinoreupha-8-ene-24-oate(1)、iso-maticadienediol(2)、25,26,27-trinortirucall-8-ene-3β-ol-24-acid(3)、dammarendiolⅡ(4)、eupha-8,24-diene-3-ol-26-al(5)、lnonotusane C(6)、eupha-8,24-diene-3β-ol-7,11-dione(7)、inoterpene A(8)、inoterpene B(9)和eupha-24-methylene-8-ene-3β-ol-7,11-dione(10),其中化合物1为新天然产物,化合物2~4首次从该科植物中分离得到,化合物5和6首次从该属植物中分离得到。细胞毒活性测试结果显示化合物1~10对MCF-7,U937以及C6细胞株均未表现出明显的细胞毒活性。     

钩毛茜草的化学成分及其细胞毒活性研究

综合采用多种色谱分离技术对钩毛茜草Rubia oncotricha根及根茎的甲醇提取物进行化学成分研究,结合NMR、MS等波谱技术,共分离鉴定了14个化合物,分别为rubiprasin D(1)、rubiprasin B(2)、rubiprasin C(3)、齐墩果酸(oleanolic acid,4)、methyl-5-hydroxy-dinaphtho[1,2-2’3’]furan-7,12-dione-6-carboxylate(5)、rubioncolin C(6)、大叶茜草素(mollugin,7)、呋喃大叶茜草素(furomollugin,8)、3-氨基-2-甲氧羰基-1,4-萘醌(3-amino-2-methoxycarbonyl-1,4-naphthoquinone,9)、1-羟基-2-甲基蒽醌(1-hydroxy-2-methyl-9,10-anthraquinone,10)、2-羟基-6-甲基蒽醌(2-hydroxy-6-methyl-9,10-anthraquinone,11)、1,4-二羟基-2-羟甲基蒽醌(1,4-dihydroxy-2-hydroxymethyl-9,10-anthraquinone,12)、2-羟基-1-甲氧基蒽醌(2-hydroxy-1-methoxy-9,10-anthraquinone,13)、1-羟基-2-甲氧基-6-甲基蒽醌(1-hydroxy-2-methoxy-6-methyl-9,10-anthraquinone,14)。化合物类型涉及萜类和醌类,其中化合物1为新的齐墩果烷型三萜类化合物,化合物25,9,1113为首次从该植物中分离得到。对所有化合物进行细胞毒和杀线虫活性筛选,结果显示化合物4,6,11,12对人非小细胞肺癌细胞(A549),人胃癌细胞株(SGC-7901)和人宫颈癌细胞(He La)具有一定的生长抑制作用,以6活性最强,其IC50分别为19.42,2.74,8.07μmol·L^-1。     

Anti-Proliferative Activity of Triterpenes Isolated from Clinicanthus nutans on Hep-G2 Liver Cancer Cells

Problem statement: Clinicanthus nutans has been used by Malaysian since long time ago. It is used to treat manydiseases including cancer. Many studies carried out on its crude extract but no clear report on the specific secondarymetabolites responsible for its nature in treating selected diseases. Objective: This study aims to confirm the practicecarried out by many people on the usage of Clinicanthus nutans in treating cancer. Methods: C. nutans leaves wereextracted by methanol. Thin layer chromatography was used to identify the suitable solvent for fractions separation.The fractions were then separated at larger volume using gravity column chromatography. Each fraction was tested onits anti-proliferative activity on Hep-G2 liver cancer cells by MTT assay. The phytochemical screening was carried outto identify the bioactive compound based on qualitative analysis. Results: The fraction 2 (F2) of C. nutans showed thelowest IC50 value of 1.73 μg/ml against Hep-G2 cancer cells, and it is identified as triterpenes. Conclusion: The fractionF2 identified as triterpenes isolated from C. nutans has potential as an anti-proliferative agent against liver cancer.     

灵芝三萜对人血单核细胞白血病细胞株THP-1的作用

目的 探讨灵芝三萜对急性单核细胞白血病细胞株THP-1作用及其可能的机制.方法 采用不同水平灵芝三萜作用于THP-1细胞48小时后,噻唑兰(MTT)比色法测定细胞增殖抑制率;流式细胞术(FCM)检测细胞周期分布;钙依赖磷脂结合蛋白异硫氰酸荧光素/碘化丙啶(annexin V-FITC/PI)双染法流式细胞仪检测凋亡率的变化.结果 MTT测定细胞增殖抑制率,与灵芝三萜0 mg/L比较,灵芝三萜80、120、160、200 mg/L抑制THP-1细胞增殖,抑制率逐渐升高(P＜0.01),分别为(21.58±3.15)％、(54.36±4.49)％、(62.10±4.23)％和(70.20±4.21)％,呈剂量依赖性.Annexin V-FITC/PI双染显示,80、120、160、200 mg/L的灵芝三萜作用THP-1细胞48小时后产生诱导凋亡作用,早期凋亡率分别为(18.6±4.30)％、(28.1±5.88)％、(40.46±6.39)％、(54.22±2.00)％,随着灵芝三萜浓度的增加有增加趋势(P＜0.01).FCM检测细胞周期,80、120、160、200 mg/L的灵芝三萜作用THP-1细胞48小时后,G2/M期、S期细胞数明显减少,G0/G1期细胞数明显增加,与对照组比较差异有统计学意义(P＜0.01).结论 灵芝三萜有抑制THP-1细胞增殖的作用,其可能的机制是阻止细胞周期在抑制G1期和诱导细胞凋亡.     

Anti‐diabetic effect of standardized extract of Potentilla discolor Bunge and identification of its active components

Potentilla discolor Bunge, a Chinese folk medicine, has been used for the treatment of diabetes for many years. Unfortunately, little information is available on its bioactive components and mechanism of action. We report the anti‐diabetic effect of P. discolor on normal and alloxan‐induced diabetic mice, as well as the possible constituents and mechanism responsible for this activity. We found that the standardized extract of P. discolor (EPD) had little effect on the glucose levels of normal mice, while a dose‐dependent hypoglycemic effect was observed in diabetic mice. Glucose tolerance test data indicated that there was a significantly higher rate of glucose disposal after EPD treatment. Phytochemical characterization indicated that the major components of EPD were triterpenes and flavonoids that could inhibit the activity of glycogen phosphorylase (GP) in vitro. Tormentic acid, asiatic acid, and potengriffioside A exhibited inhibitory effects on GP (IC₅₀=90.6, 65.4, and 85.9 µM, respectively). The inhibition on GP may be one molecular mechanism through which EPD ameliorates hyperglycemia. These results suggest P. discolor extract may offer an alternate treatment for diabetes. Drug Dev Res, 2009. © 2009 Wiley‐Liss, Inc.