鹰嘴豆中3种异黄酮及其乙氧基化衍生物体外协同降糖活性研究＊

目的：通过对鹰嘴豆中3种异黄酮类化合物染料木素、鹰嘴豆芽素A、刺芒柄花素进行乙氧基化结构修饰,研究其降糖活性及协同降糖活性。方法：对上述3种异黄酮类化合物进行乙氧基化结构修饰,并研究该3种异黄酮类化合物及其衍生物降糖活性,并对化合物进行组合给药,研究其之间存在的协同降糖活性,选择胰岛素抵抗HepG2细胞作为降糖活性筛选模型。结果：获得4个乙氧基化产物,3个母体异黄酮衍生物中染料木素的降糖活性优于鹰嘴豆芽素A和刺芒柄花素,差异具有统计学意义（P<0.05）,3个母体异黄酮衍生物最大给药剂量与阳性药盐酸二甲双胍相比,效果不如盐酸二甲双胍,差异均具有统计学意义（P<0.05或P<0.01）。衍生物中化合物a和d之间差异无统计学意义（P>0.05）,化合物b和c降糖效果均不如化合物a和d,差异具有统计学意义（P<0.05）,其中化合物d较修饰前降糖活性有明显提高（P<0.05）。组合5的降糖活性与盐酸二甲双胍相当,差异无统计学意义（P>0.05）。结论：本研究发现,通过化合物组合,化合物之间可以起到协同作用,发挥更好的降糖效果,为开发具有自主知识产权的降糖药物提供一定的基础。     

鹰嘴豆中三种异黄酮酯化反应研究

目的:改善鹰嘴豆中3种异黄酮的药物活性。方法:以鹰嘴豆芽素A、刺芒柄花素、染料木素为原料,对其进行4-溴丁酰酯化反应、EDC缩合反应、正丁酰酯化反应。结果:合成了12个异黄酮类衍生物,方法简单、条件易控、产率较高。结论:产物结构通过1H-NMR、13C-NMR、ESI-MS分析确认,为进一步对其药物活性研究奠定结构基础。     

鹰嘴豆中三种异黄酮醚化反应研究

目的：改善鹰嘴豆中三种异黄酮的药物活性。方法：以鹰嘴豆芽素A、刺芒柄花素、染料木素为原料,以丙酮为溶剂,碳酸钾作为催化剂,分别与1,3-二溴丙烷,1-溴丙烷,3-溴丙烯进行醚化反应。结果：合成了9个异黄酮类衍生物,方法简便、条件易控、产率较高。结论：产物结构通过1H-NMR、13C-NMR、ESI-MS分析确认,为进一步对其生物活性研究奠定结构基础。     

鹰嘴豆中三种异黄酮结构修饰衍生物的合成及其结构表征

目的:改善鹰嘴豆中三种异黄酮的溶解性,提高其生物利用度。方法:以刺芒柄花素、鹰嘴豆芽素A、染料木素为原料,对其进行异丁基化反应、异丁酰酯化反应、苯磺酰酯化反应。结果:合成了10个异黄酮类衍生物,方法简便、条件易控、产率较高。结论:产物结构通过1H-NMR、13C-NMR、ESI-MS分析确认,为进一步对其生物活性研究奠定结构基础。     

一测多评法测定红车轴草中6种异黄酮类成分含量

目的:建立红车轴草中大豆黄素、染料木素、红车轴草素、芒柄花素、樱黄素和鹰嘴豆芽素A的一测多评方法。方法:采用HPLC法,以鹰嘴豆芽素A为内参物,分别建立大豆黄素、染料木素、红车轴草素、芒柄花素和樱黄素的相对校正因子,并计算6种异黄酮类成分的含量。同时采用外标法测定6种成分的含量,并比较两种测定方法间的差异性。结果:一测多评法与外标法测定结果无显著差异,各相对校正因子重复性良好。结论:一测多评测定红车轴草中6种异黄酮类成分含量的方法准确可靠,可用于红车轴草药材的质量控制。     

鹰嘴豆化学成分及药理作用研究进展

通过对国内外近年来关于鹰嘴豆化学成分及药理作用的相关文献进行归纳和总结,对鹰嘴豆的化学成分及药理作用进行了综述,为今后进行鹰嘴豆化学成分的研究及活性成分的发现提供基础。通过文献研究发现,鹰嘴豆含有多种营养素及人体必需物质如葡聚糖、蔗糖和鹰嘴豆糖以及阿糖配葡聚糖,胆碱磷脂,各类维生素、氨基酸以及泛酸和植酸,并含有各类微量元素如磷、钙、铁、锌等,以及人体不能自行合成的不饱和脂肪酸。其目前研究较多的具有活性的成分主要为三萜皂苷类成分及异黄酮等,其中目前已经获得的三萜皂苷类主要有大豆精醇A,β-香树脂醇,β-谷甾醇,大豆皂苷Bb以及胡萝卜苷等,异黄酮类化合物主要有鹰嘴豆芽素A、芒柄花素、染料木素、毛蕊异黄酮、芒柄花苷、红车轴草苷、印度黄檀苷。这些三萜皂苷及异黄酮组分具有调节血糖、降低血脂等诸多药理作用。另外,鹰嘴豆及其主要活性成分还具有降胆固醇、抑制CaCo-2细胞、改善学习记忆能力、抑制肿瘤细胞的生长等作用。鹰嘴豆在不同的生长阶段其含有的各类物质也在不断的发生着变化,对于不同的疾病也起到不同的治疗效果,随着鹰嘴豆研究的不断深入,将会发现其中更多的化学成分,并且其中活性成分也将逐渐的被筛选出来,将大大扩展鹰嘴豆的应用范围。     

补阳还五汤中异黄酮和紫檀烷类化合物的分离和结构鉴定

目的 研究补阳还五汤的抗血栓有效部位及其活性成分。方法 用多种色谱和光谱方法对补阳还五汤水煎液乙酸乙酯萃取部位的化学成分进行分离和结构鉴定。结果 分离鉴定了4个异黄酮化合物和1个紫檀烷类化合物,分别为芒柄花素（Ⅰ）、毛蕊异黄酮（Ⅱ）、芒柄花素－7－O－β－D－葡萄吡喃糖苷（Ⅲ）、毛蕊异黄酮－7－O－β－D－葡萄吡喃糖苷（Ⅳ）及（6aR,11aR)9,10-二甲氧基紫檀烷－3－O－β－D－葡萄吡喃糖苷（Ⅴ）。结论 以上化合物均为首次从补阳还五汤中分离得到。     

芒柄花素抗肿瘤作用机制的研究进展

芒柄花素（FN）是一种从红三叶草、黄芪和鸡血藤等药用草本植物中提取的植物异黄酮。研究表明,芒柄花素具有较强的抗肿瘤生物活性,可作为抗肿瘤药物治疗各种恶性肿瘤。迄今为止的研究表明,芒柄花素通过多种分子机制与途径对肿瘤产生抑制增殖、诱导凋亡、抑制迁移和侵袭、诱导细胞周期停滞等作用,可以在乳腺癌、结直肠癌、前列腺癌、膀胱癌和肺癌等多种肿瘤细胞和动物模型中观察到这些抗肿瘤活性。主要抗肿瘤活性体现在触发活性氧（ROS）生成、调节磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/雷帕霉素靶蛋白（mTOR）和丝裂原活化蛋白激酶（MAPK）信号通路;抑制酪氨酸激酶1（JAK1）,酪氨酸激酶2（JAK2）和非受体酪氨酸激酶（c-Src）的激活;调节细胞角蛋白19（CK19）,基质金属蛋白酶（MMP）,微小RNA-21（miR-21）,核纤层蛋白A/C抗体（Lamin A/C）,细胞周期蛋白D₁（Cyclin D₁）和细胞周期蛋白E₁（Cyclin E₁）的表达。此外,笔者对于芒柄花素衍生物的抗肿瘤作用研究也进行了综述。通过对芒柄花素的化学结构进行修饰,目前已得到多种具有抗肿瘤作用的衍生物。实验结果表明芒柄花素的一些衍生物具有更强的抗肿瘤活性和较低的细胞毒性,分子作用机制研究仍需要进一步深入。综上所述,芒柄花素及其衍生物可能成为抗肿瘤的潜在新药。     

异黄酮对活性氧引起脂质过氧化的抑制作用

对于类黄酮有清除活性氧及抑制脂质过氧化的作用的报道较多,但未进行详细探讨。本次探讨了与其作用相关的异黄酮(鹰嘴豆芽素A、大豆黄素、刺芒柄花素、金雀异黄素)对活性氧引起脂质过氧化的抑制作用。     

红车轴草化学成分的研究

 目的对红车轴草(Trifolium pratense L.)的化学成分进行研究。方法应用硅胶柱层析和Sephadex LH-20凝胶柱层析对化合物进行分离,通过理化和波谱分析方法鉴定其结构。结果从红车轴草中分离得到8个化合物,分别鉴定为十八烷醇(Ⅰ)、5-羟基-7,4′-二甲氧基异黄酮(Ⅱ)、鹰嘴豆芽素A(Ⅲ)、樱黄素(Ⅳ)、芒柄花素(Ⅴ)、染料木素(Ⅵ)、芒柄花苷(Ⅶ)和大豆苷元(Ⅷ)。结论化合物Ⅰ和Ⅱ为首次从该植物中得到。     

Inhibitory effects of isoflavones on lipid peroxidation by reactive oxygen species

Possible inhibitory effects were investigated for four isoflavones, biochanin A, daidzein, formononetin and genistein on lipid peroxidation by reactive oxygen species. Biochanin A, formononetin and genistein inhibited lecithin peroxidation which was induced by hydroxy radical generation, by interaction of haemoglobin and hydrogen peroxide. Daidzein and formononetin inhibited lecithin peroxidation which was induced by superoxide anion generation by xanthine-xanthine oxidase. These results demonstrated that the differences in antioxidant activities of isoflavones are dependent on the relation between their chemical structures and reactive oxygen species.     

Antiplasmodial activity of isoflavones from Andira inermis

The stem bark and seeds of Andira inermis, Fabaceae, are employed as a purgative, vermifuge, and febrifuge. In particular, the powdered bark is claimed to be efficacious in intermittent fever. Bioassay-guided fractionation of lipophilic extracts from the stems and leaves yielded six isoflavones: biochanin A, calycosin, formononetin, genistein, pratensein, and prunetin. Calycosin (3', 7-dihydroxy-4'-methoxyisoflavone) and genistein (4',5, 7-trihydroxyisoflavone) have been shown to possess in vitro activity against the chloroquine-sensitive strain poW and the chloroquine-resistant clone Dd2 of Plasmodium falciparum.     

Inhibitory Effects of Isoflavonoids on Rat Prostate Testosterone 5α-Reductase

Testosterone 5α-reductase inhibitors represent important therapeutic drugs for use against androgen-dependent diseases such as benign prostatic hyperplasia, male pattern baldness, and acne. We have searched for inhibitors of rat prostate testosterone 5α-reductase in the cultured broths of many kinds of soil bacteria, and have found that cultured soybean-casein digest broths of certain bacterial strains have a potent inhibitory effect on the enzyme. We tested 10 selected isoflavonoids, including isoflavones and O-methylated isoflavones, for inhibitory effects on rat prostate testosterone 5α-reductase to determine the important structural elements for inhibition of the enzyme. Genistein, biochanin A, equol, and 3′,4′,7-trihydroxyisoflavone showed considerably higher inhibitory effects whereas daidzein, formononetin, glycitein, prunetin, ipriflavone, and 4′,7-dimethoxyisoflavone showed lower inhibitory effects. The IC₅₀ values of genistein, biochanin A, equol, 3′,4′,7-trihydroxyisoflavone, and riboflavin, a positive control, for rat prostate testosterone 5α-reductase were 710 μm, 140 μm, 370 μm, 690 μm, and 17 μm, respectively. Daidzein, genistein, biochanin A, formononetin, and equol are already known to be testosterone 5α-reductase inhibitors, but this is the first characterization of 3′,4′,7-trihydroxyisoflavone as an inhibitor of the enzyme.     

Antigiardial activity of isoflavones from Dalbergia frutescens bark

Several isoflavones [formononetin (1), castanin (5), odoratin (6), glycitein (7), pseudobaptogenin (8), fujikinetin (9), and cuneatin (10)] were isolated from Dalbergia frutescens, and their antiprotozoal activities were determined against Giardia intestinalis. Among these compounds, formononetin (1) was the most potent antigiardial agent, with an IC(50) value of 30 ng/mL (approximately 0.1 microM), as compared to the value for metronidazole, the current drug of choice, of 100 ng/mL (approximately 0.6 microM). Three isoflavones closely related to formononetin [daidzein (2), biochanin A (3) and genistein (4)] were also evaluated, but they were at least 100 times less active than 1. Formononetin (1) may thus be an interesting lead for development of new antigiardial agents or as a probe for a new mechanistic target.     

Isoflavones as apoptosis inducers in human hepatoma HuH-7 cells

Nine flavonoids isolated from the ethyl acetate extract of Pycnanthus angolensis were assayed for their potential apoptosis induction activities in human hepatoma HuH-7 cells. These flavonoids include eight isoflavones, namely irilone (1), tectorigenine (2), formononetin (3), genistein (4), 2'-hydroxybiochanin A (5), mixture of biochanin A (6) and prunetin (7), and 4',7-dihydroxy-2'-methoxyisoflavan (8), and the flavanone liguiritigentin (9). Their chemical structures were characterized by spectroscopic methods including 2D NMR experiments. Methodology for cell death detection included the LDH assay, Hoechst staining, TUNEL staining and general caspase-3-like activity assay. The compounds tested showed higher apoptosis induction profiles in HuH-7 cells compared with the control. Caspase activity assays confirmed the apoptosis inducing activity of these flavonoids.     

The isoflavones mixture from Trifolium pratense L. protects HCN 1-A neurons from oxidative stress

Oxidative stress-induced neuronal cell death has been implicated in different neurological disorders and neurodegenerative diseases such as Alzheimer's disease and Parkinson's. Using the Alzheimer's disease-associated hydrogen peroxide (H(2)O(2)), we investigated the neuroprotective efficacy of a natural mixture of phytoestrogenic isoflavones (genistein, daidzein, biochanin A and formononetin) from Trifolium pratense L. (Red clover) against oxidative stress-induced cell death in human cortical cell line HCN 1-A maintained in culture. Neuronal viability was determined by MTT or trypan blue test and neuronal integrity by morphological analysis.The results obtained indicate that exposure of HCN 1-A cell cultures to hydrogen peroxide resulted in a concentration-dependent decrease in neuron viability. Concentration of H(2)O(2) ranging from 50 to 200 microg/ml were toxic to these cultures. A 24-hour pretreatment with 0.5, 1 and 2 microg/ml isoflavones extract significantly increased cell survival as evidenced by MTT or trypan blue test and significantly prevented the morphological disruption caused by H(2)O(2) as shown by microscopical inspection, indicating that neurons treated with isoflavones were protected from the cell death induced by H(2)O(2) exposure. These findings imply that the neuroprotective effect of isoflavones extract is partly associated with its antioxidant activity. Further, results of these investigations indicate that although isoflavones extract exert a neuroprotective effect, it do not promoted cortical neuron process outgrowth.     

丰城鸡血藤异黄酮类成分的研究

目的：研究丰城鸡血藤的化学成分。方法：用色谱法分离,用理化性质及波谱方法鉴定结构。结果：从中分离鉴定了4个异黄酮类化合物：3′-O-methylorobol(1),染料木素(genistein,2),鹰嘴豆芽素A(biochanin A,3),阿弗洛莫生(afromosin,4)。结论：4个化合物均为首次从该种植物中分离得到。     

苦瓜蛋白酶抑制剂对苦瓜降糖多肽活性的保护作用

目的 探讨苦瓜蛋白酶抑制剂对苦瓜降糖多肽口服降糖活性的保护作用.方法 单次iv四氧嘧啶80mg/kg给药建立糖尿病小鼠模型.以降糖多肽(20、10、5 mg/kg)iv给药.于0.5、2、4、6、8 h尾静脉取血测定空腹血糖;测定在不同降糖多肽与蛋白酶抑制剂配伍剂量下,ig给药21 d后的空腹血糖值;以降糖作用显著的配伍比例,ig给药7、14、21 d和停止给药7 d后测定血糖.结果 降糖多肽于iv给药4 h后,高、中、低3个剂量均表现出显著降血糖效应(P<0.05、0.01);降糖多肽与抑制剂(300+300)、(100+100)、(100+50)、(100+25)mg/kg 4个配伍剂最组ig给药21 d后表现出显著降血糖效应,且以后两个剂量组ig给药的降血糖作用为更佳,停药7 d后给药组血糖值与模型组无显著差异.结论 苦瓜蛋白酶抑制剂在ig给药过程中能够使苦瓜降糖多肽免遭消化系统的蛋白水解酶破坏而起保护作用,维持其降血糖药效,该作用与两者间的配伍剂量有关.