18 resultados para Perseus, Medusa, beheading


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水母雪莲(Saussurea medusa Maxim.)为多年生菊科植物,是我国珍稀药用资源。所含的主要生物活性成分是黄酮类物质,具有抗炎、镇痛、免疫抑制及抗氧化等功效。但由于水母雪莲生长环境特殊,生长缓慢,人工引种困难;加上长期掠夺性采挖,造成其野生药用资源短缺,已经不能满足市场的需求。近年来,世界上掀起了植物药开发的热潮,植物药以其天然低毒的特点倍受关注,而黄酮类化合物更是以其广谱的药理作用引人瞩目。 黄酮类化合物的合成代谢途径在植物界进化过程中很保守,黄酮类生物合成途径中的相关酶也已得到确证并进行了系统的研究。二氢黄酮醇-4-还原酶(Dihydroflavonol-4-reductase, DFR)是一个处于花色素或者原花色素合成途径中的关键酶,它与黄酮合成途径中的黄酮醇合成酶(flavonol synthase)竞争底物。本研究以水母雪莲为研究对象,根据近缘物种DFR基因的保守核苷酸序列设计兼并引物,通过PCR技术,从已经建立的水母雪莲红色愈伤组织cDNA文库中筛选到一个编码该酶的cDNA序列,该序列全长1166个碱基对。根据生物信息学分析,此cDNA编码342个氨基酸;Blastp分析结果显示,该氨基酸序列与同科植物翠菊(Callistephus chinensis)的相似性最高,达87%;SWISSMODEL软件预测其蛋白的三级结构与葡萄(Vitis vinifera)的十分相似,活性中心的关键氨基酸残基也完全一致。据此可以断定,我们所得到的cDNA为编码二氢黄酮醇还原酶的基因,并命名为水母雪莲二氢黄酮醇还原酶基因(SmDFR)。为了得到SmDFR的DNA序列,我们又设计特异引物,从水母雪莲的基因组中扩增出了由1871个碱基对组成的DNA序列,该序列包含五个内含子和六个外显子。 为了提高水母雪莲和大苞雪莲中黄酮类物质的含量,我们构建了SmDFR的反义植物表达载体,利用根癌农杆菌介导进行基因转化。通过改变影响农杆菌转化的实验条件包括外植体来源、农杆菌菌株、细菌浓度、外植体预培养时间、侵染时间和乙酰丁香酮的浓度进行转基因试验,目前尚未得到转基因植株。另外,我们构建了SmDFR正义植物表达载体,通过对拟南芥(Arabidopsis thaliana) DFR基因突变体和矮牵牛(Petunia hybrida)进行基因转化,来验证SmDFR的功能;目前,此实验尚在进行之中。

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本学位论文报道了作为传统藏药材广泛使用的西藏产雪莲花化学成分的研究。论文由五章组成,第一章是三种西藏产雪莲花的化学成分的系统分离纯化和结构鉴定;第二章为西藏产雪莲花化学成分的液-质及串联质谱联用分析;第三章提出了以HPLC和TLC为检测方法的雪莲花药材质量标准草案;第四章给出了对西藏产雪莲花挥发油化学成分的气-质联用分析结果;第五章概述了雪莲花的化学成分及药理研究进展。 第一章包括三个部分。第一部分报道了绵头雪莲花(Saussurea laniceps Hand.-Mazz.)全草乙醇提取物化学成分的分离鉴定。采用正相硅胶柱层析及凝胶柱层析等分离方法,从西藏产绵头雪莲花的乙醇提取物中共分离鉴定出15个化合物。其中11个化合物为首次从该植物中分离得到,当中2个化合物系在凤毛菊属植物中首次发现。第二部分报道了水母雪莲花(Saussurea medusa Maxim.)全草乙醇提取物的化学成分。采用正、反相硅胶柱层析及凝胶柱层析等分离方法,共分离鉴定出15个化合物,其中1个为新化合物,另有4个化合物为首次从该植物中分离得到。新化合物结构通过质谱和一维及二维核磁共振等波谱解析方法及碱水解反应确定为巴豆酰基-高车前苷(M-7)。第三部分报道了三指雪莲花 (Saussurea tridactyla Sch.-Bip. ex Hook. f.)全草乙醇提取物的化学成分。采用正相硅胶柱层析及凝胶柱层析等分离方法,共分离鉴定出7个化合物,其中1个化合物为首次从该植物中分离得到。 第二章也包括三个部分。首先是采用液-质联用(HPLC-DAD-ESI-MSn)分析方法,对7个西藏不同产地的三指雪莲花化学成分进行了分析,通过与标准品的 UV和MS数据比较,共鉴定出14个峰,并对其中8个共有成分进行了定量测定。其次是关于八种西藏产雪莲花化学成分的液-质联用(HPLC-DAD-ESI-MSn)分析,通过与标准品的UV和MS数据比较,共鉴定出15个峰,并对其中8个共有成分进行了定量检测。最后通过对八种西藏产雪莲花主要化学成分的多级串联质谱(ESI-MSn)分析,快速、灵敏地鉴定出10个黄酮和3个香豆素化学成分。 第三章同样包括三个部分。首先是以绵头雪莲花中主要香豆素成分东莨菪素和伞形花内酯为对照品,通过TLC定性检测和HPLC含量测定,草拟出较严谨的药材质量标准。其次是将绵头雪莲花、三指雪莲花和雪兔子作为一个药材看待,草拟了以东莨菪素和伞形花内酯的TLC检测为指标的药材质量标准。最后是针对水母雪莲花,以主要黄酮成分芹菜素-7-O-b-D-葡萄糖苷为对照品作TLC检测,并草拟出该药材的质量标准草案。 第四章报道了西藏产雪莲花挥发油的化学成分分析。采用传统水蒸气蒸馏法分别从八种雪莲花全草中提取挥发油,利用气相色谱-质谱联用技术分别从水母雪莲花、绵头雪莲花、槲叶雪莲花、云状雪兔子、拉萨雪兔子、小果雪兔子、雪兔子和三指雪莲花中分别鉴定出83、83、56、34、21、20、24和20个化学成分,分别占其挥发油总量的70.7%、76.0%、82.2%、55.4%、49.7%、70.4 %、76.2%和 76.7%。 第五章为综述,总结和概括了雪莲花的化学和药理研究进展。 The dissertation reports the investigation of the chemical constituents of the genus Saussurea. Quite a lot of species in this genus are traditional Tibetan medicinal plants, and hence have been widely used in traditional Tibetan medicine. This dissertation consisted of five chapters. The first chapter is on the chemical constituents of three Saussurea plants. The second section is about the analysis of chemical constituents of Saussurea plants using HPLC-MS and ESI-MS/MS. In the third chapter, we proposed quality-control standards for the Genus Saussurea based on TLC (thin layer chromatography) and HPLC. The fourth chapter is about chemical compositions of the essential oil from the whole plant of Saussurea plants. The last chapter reviews the research progress of the Genus Saussurea. The first chapter consists of three parts. The first part is about chemical constituents of ethanol extracts from whole plant of Saussurea laniceps Hand.-Mazz. Fifteen compounds were isolated by column chromatography on normal phase silica gel and Sephadex LH-20. Among them, eleven compounds were isolated from this plant for the first time, and two compounds were isolated from Genus Saussurea for the first time. The second part is about chemical constituents of ethanol extracts from whole plant of Saussurea medusa Maxim. Fifteen compounds were isolated by column chromatography on normal phase, reversed phase silica gel and Sephadex LH-20. Five of them were isolated from this plant for the first time, and there is one new flavonoid glucoside which was identified as 6″-O-crotonoyl-homoplantaginin (M-7) based on the evidence of one- and two-dimensional nuclear magnetic resonance, mass spectrometry analysis, and alkaline hydrolysis reaction. The last part is about chemical constituents of ethanol extracts from whole plant of Saussurea tridactyla Sch.-Bip. ex Hook. f.. Seven compounds were isolated by column chromatography on normal phase silica gel and Sephadex LH-20. There is one compound which was isolated from this plant for the first time. The second chapter consists of three parts. In the first part, we analyzed the chemical constituents of S. tridactyla collected from seven different places in Tibet using HPLC-DAD-ESI-MSn. Fourteen peaks in the HPLC were identified by comparison of UV and MS spectra with those of authentic compounds, among which eight common peaks were quantified. In the second part, we analyzed the chemical constituents of eight Saussurea species using HPLC-DAD-ESI-MSn method. Fifteen peaks in the HPLC were identified by comparison of UV and MS spectra with those of authentic compounds and eight main peaks of them were quantified. In the last part, we analyzed the chemical compounds of the above eight Saussurea plants directly by ESI-MS/MS. Thirteen major compounds, including 10 flavonoids and 3 coumarins were easily rapidly identified. The third chapter consists of three parts. In the first part, we proposed a comparative high quality-control standard for S. laniceps, based on quality detection by TLC and quantity analysis by HPLC using two major compounds (umbelliferone and scopoletin) as standard compounds. In the second part, in viewing S. laniceps, S. tridactyla and S. gossypiphora as the members of one family of medicinal herbs, we suggested a quality-control standard based on the TLC detection of the two major compounds (umbelliferone and scopoletin). In the last part, we proposed a quality-control standard for S. medusa based on the TLC detection of its major component (apigenin 7-O-glucoside). The four chapter analyzed the chemical constituents of essential oil of eight Saussurea species. The essential oils were extracted from the whole plants of these samples with water stream distillation. By GC-MS analysis, we identified eighty-three compounds from S. medusa, eighty-three from S. laniceps, fifty-six from S. quercifolia, thirty-four from S. aster, twenty-one from S. kingii, twenty from S. simpsoniana, twenty-four from S. gossypiphora, and twenty from S. tridactyla respetively, which accounted for 70.7%, 76.0%, 82.2%, 55.4%, 49.7%, 70.4 %, 76.2% and 76.7% of the total essential oil, respectively. The last chapter reviews the research progress of the Genus Saussurea.

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以水母雪链(Saussurea medusa Maxin.)无菌幼苗的根作外植体,诱导获得的愈伤组织为材料,研究光照对生物碱合成的影响。结果表明:尽管在水线雪链的愈伤组织培养物中,总生物碱的含量比原植株中的含量低,但其组成成分比原植物丰富,并且秋水仙碱的含量也比原植物高,尤其是在光照培养的愈伤组织中秋水仙碱大大增加。