双花千里光、川芎及宽叶羌活的化学成分研究

本论文由四章组成。第一、二和三章分别报道了双花千里光、川芎和宽叶羌活的化学成分研究。从三种药用植物中共分离和鉴定了40 个化学成分，其中8个为新化合物。第四章概述了藳本属植物及日本川芎的化学成分研究进展。 第一章包括三个部分。第一部分报道双花千里光（Senecio dianthus Franch.）地上部分乙醇提取物的化学成分。采用正、反相硅胶柱层析等各种分离方法，从中共分离出8 个艾里莫酚型倍半萜内酯，其中5 个是新化合物，并且有1 个为首次发现的连接了含氮原子取代基的艾里莫酚型倍半萜内酯。它们的结构经MS、IR、NMR及X-单晶衍射等解析方法确定为2b-angeloyloxy-10b-hydroxyeremophil-7(11)-en-8a,12-olide (1)、6b-angeloyloxy-10b-hydroxyeremophil-7(11)-en-8a,12-olide (2)、2b-angeloyloxy-8b,10b- dihydroxyeremophil-7(11)-en-8a,12-olide (3)、2b-angeloyloxy-8a-hydroxyeremophil-7(11),9(10)-dien-8b,12-olide (4)和8b-amino-10b- hydroxyleremophil-7(11)-en-8a,12-olide (5)。这8 个倍半萜内酯经体外生物活性测试表明均具有通过抑制巨噬细胞增殖抵制破骨细胞增生的活性。第二部分对艾里莫酚型倍半萜内酯的质谱裂解规律进行了初步探讨。第三部分报道双花千里光茎、和叶花的挥发油成分分析。采用传统水蒸气蒸馏法分别提取了双花千里光茎、叶和花的挥发油，用气相色谱－质谱联用（GC-MS）技术分别分离鉴定了其化学成分，从茎、叶和花挥发油中各分离和鉴定出70、80 和73 种化学成分，分别占挥发油总量的91.2％、85.7％及93.4％。 第二章包括两个部分。第一部分报道川芎（Ligusticum chuanxiong Hort.）根茎乙醇提取物的化学成分。通过正、反相硅胶柱层析等分离纯化和MS、NMR及X-单晶衍射等解析方法，共分离鉴定了21 个化合物，结构类型分属于苯酞、二聚苯酞、香豆素和脂肪酸类。其中2 个为结构比较新颖的二聚苯酞类化合物：chuanxiongnolide A (19)和chuanxiongnolide B (20)，化合物19 的结构经X-单晶衍射得到确证。第二部分报道川芎挥发油的化学成分。采用不同的提取方法（溶剂萃取法、水蒸气蒸馏法、CO2 超临界流体萃取法）提取川芎挥发油，同时采集不同产地（四川彭县、四川郫县、云南鹤庆）及不同品质（川芎、奶芎、苓子）的川芎产品，利用GC-MS 技术分离鉴定其挥发油的化学成分，计算各成分的相对含量，并对比分析其中的异同。 第三章报道宽叶羌活（Notopterygium forbesii Boiss.）根茎化学成分的分离纯化和结构鉴定。通过正、反相硅胶柱层析等分离纯化和MS、NMR 等解析方法，共分离鉴定了13 个化合物，结构类型分属于香豆素、二氢异香豆素、甾体和羧酸类。其中1 个新二氢异香豆素类成分鉴定为6-methoxy-hydrangenol (37)。 第四章概述了藳本属植物及日本川芎化学成分的研究进展。 This dissertation consisted of four chapters. The former three chaptersrespectively elaborated the phytochemical investigation of three herbal medicines:Senecio dianthus Franch., Ligusticum chuanxiong Hort. and Notopterygium forbesiiBoiss.. Forty compounds, including eight new ones, were isolated and identified byspectral and chemical evidence. The fourth chapter elaborated the study progress ofchemical constituents of Ligusticum genus and Cnidium offcinale. The first chapter consisted of three parts. The first part is about the chemicalconstituents of ethanol extraction and essential oils from the aerial parts of S. dianthu.Eight eremophilenolides were isolated and identified. Among them, five ones are newcompounds and one of them is a novel eremophilenolide attched with an amino group.The structures of the new compounds were identified as 2b-angeloyloxy-10b-hydroxyeremophil-7(11)-en-8a,12-olide (1)，6b-angeloyloxy-10b-hydroxyeremophil-7(11)-en-8a,12-olide (2)，2b-angeloyloxy-8b,10b-dihydroxyeremophil-7(11)-en-8a,12-olide (3)，2b-angeloyloxy-8a-hydroxyeremophil-7(11),9(10)-dien-8b,12-olide (4) and8b-amino-10b-hydroxyeremophil-7(11)-en-8a,12-olide (5) by spectral evidence andX-ray crystallography analysis. All the compounds were evaluated for theiranti-osteoclstogenesis activity using a proliferation inhibit assay with microphagecells. The second part elementarily discussed the characteristic fragmentation oferemophilenolides isolated from S. dianthus in ESI-MS.The latter part is about thechemical constituents of essential oil extracted from stems, leaves and flowers of S.dianthus with steam distillation. By the GC-MS analysis, 70, 80 and 73 compoundswere respectively isolated and identified which accounted for more than 91.2％, 85.7％ and 93.4％ of total essential oil. The second chapter, including two parts, is about the the chemical constituents ofethanol extraction and essential oils from rhizomes of L. chuanxion. In the first part, twenty-one compounds were isolated and iedntified. Two ones are novel dimericphthalides and the structures were suggested as chuanxiongnolide A (19) andchuanxiongnolide B (20) by spectral evidence and confirmed by X-raycrystallography analysis. In the second part, the samples were collected from differentextract techniques (solvent extraction, steam distillation and supercriticalfluid extraction), different habitats (Peng and Pi counties, Sichuan province; Heqing,Yunnan province) and different qualities (Chuanxiong, Naixiong and Lingzi). Thechemical constituents of essential oil from L. chuanxiong were analyzed by GC-MS and were compared each other. The third chapter is about the chemical constituents of rhizomas of N. forbesii,which belongs to a endemic genus of China. Thirteen compounds were isolated andidentified. One of them is a new dihydroisocoumarin and the structure was identifiedas 6-methoxy-hydrangenol (37) by spectral evidence. The fourth chapter is a review on study progress of chemical constituents ofLigusticum species and Cnidium offcinale.

中药射干的化学成分研究

本学位论文由4章组成。第一章是论文的主体，报道了中药射干的化学成分研究。第二章是中药射干代用品川射干的化学成分研究，并附带报道了西番莲化学成分的研究结果。第三章是射干、川射干及西番莲提取物化学成分串联质谱分析的报道。第四章为综述，概述了射干及鸢尾属植物的化学成分和药理研究进展。 在第一和二章中分别报道了射干（Belamcanda chinensis (L.) DC.）、川射干 （Iris tectorum Maxim.）及西番莲（Passiflora incarnate L.）化学成分的分离纯化与结构鉴定。采用正、反相硅胶柱层析、凝胶柱层析、薄层制备及HPLC等各种分离方法，从三种药用植物中共分离出68个不同的化合物，其中61个的结构得 得以鉴定，另外4个化合物的结构正在鉴定中，3个由于量少且有点杂质未作进一步的鉴定。 中药射干（Rhizoma Belamcandae）为射干植物的干燥根茎，从中共分离出53个化合物，通过红外、质谱及核磁共振等波谱方法鉴定了包括12个新化合物在内的48个，结构类型分别属于iridal型三萜及其新颖的二聚体、异黄酮、黄酮及黄酮醇、香豆素、甾体、芳香酸和脂肪酸及其甘油酯等。新化合物中有两个异黄酮类化合物，其结构分别鉴定为5,7,8,4′-四羟基-6-甲氧基异黄酮和5,6-二羟基-4′-甲氧基异黄酮-7-O-β-D-吡喃葡萄糖苷；八个新的iridal型三萜化合物分别鉴定为鸢尾烯（L）、16-甲氧基鸢尾烯、16-去羟基鸢尾烯、2-(E)-16-去羟基鸢尾烯、16-去羟基鸢尾烯B、3-乙酰基-16-去羟基鸢尾烯、iristectoroneL和iristectoroneM；两个结构骨架新颖的双三萜，分别命名为射干素A和射干素B，其分离纯化的困难以及结构的新颖和复杂突显出该论文的科学意义。除这些新化合物外，还有9个已知化合物为首次从中药射干中分离得到。此外，从中药射干的代用品川射干中分离得到7个已知化合物，主要是黄酮类成分及iridal型三萜化合物，其中1个三萜化合物为从射干中分离鉴定的新成分。另外还从西番莲中分离出8个化合物，鉴定了其中的6个，主要为黄酮碳苷。 第三章是关于射干、川射干及西番莲提取物化学成分的ESI-MS-MS分析，在初步探讨了从这些植物中分离鉴定出的一些异黄酮及黄酮碳苷的质谱裂解规律基础上，通过质谱和串联质谱分析，定性和半定量地检测了射干和川射干中主要的异黄酮成分以及西番莲中的黄酮碳苷成分，为这些药材品质的快速鉴定提供了一种简便方法。 第四章概述了射干及鸢尾属药用植物的化学和药理研究进展，特别是对其中异黄酮及三萜类成分的研究进展进行了深入系统的综述。 This dissertation is composed by four chapters. The first and second chapter reports the phytochemical investigation of three medicine plants, Belamcanda chinensis (L.)DC., Iris tectorum Maxim. and Passiflora incarnate L. Sixty eight different compounds were isolated and sixty one of them were identified. The third chapter described rapid ESI-MS-MS analysis of B. chinensis, I. tectorum, and P. incarnate. The forth part is a review about the progress of studies on the chemical constituents from Belamcanda chinensis and Iris species. Fifty-three compounds were isolated from Rhizoma Belamcandae, the rhizomes of B. chinensis by the methods of column chromatography (normal and reversed phase silica gel, Sephadex LH-20), preparative TLC and HPLC. On the basis of spectroscopic methods including IR, ESI-MS, 1-D and 2-D NMR, forty eight of them were identified as seventeen flavonoids, seventeen tritepenoids, one cumarin, five steroids and some benzene derivative etc. Among them, the structures of twelve new compounds were elucidated as 6-methoxy-5,7,8,4′-tetrahydryoxyisoflavoe, 4′-methoxy-5,6-dihydroxyisoflavone-7-O-β-D-glucopyranoside, iristectorene L, 16-methoxyisoiridogermanal, 16-dehydroxyisoiridogermanal, 2-(E)-16-dehydroxy isoiridogermanal, 16-dehydroxyiristectorene B, 3-acetyl-16-dehydroxyisoiridoger- manal, iristectorone L, iristectorone M, belamcandene A and belamcandene B. Last two new compounds are dimer of triterpenoids with a novel carbon skeleton. Beside the new compounds, nine known ones were isolated from this plant for the first time. Isolation of I. tectorum yielded seven compounds. On the basis of spectroscopic methods including ESI-MS, NMR and the comparison with authentic samples, three of them were determined as isoflavone, two of them were triterpenoids, and other two were β-sitosterol and apocynin. All of them are known compounds except one of iridal type triterpenoid, 16-dehydroxyiristectorene B, which also obtained from B. chinensis as a new compound. Isolation of P. incarnate yielded eight compounds. Six of them were determined on the basis of spectroscopic methods including ESI-MS, NMR and the comparison with authentic samples. Four of them are flavone-C-gluconside, and two are steroids. The third chapter describes the tandem mass spectrometry (ESI-MS-MS) analysis of the isoflavonoids from B. chinensis and I. tectorum, as well as C-glycosyl-flavonoide from P. incarnate, in order to explore the rapid methodology of validating the quality of the herbs. In addition, the fractionation rules of some iosflavonoids and C-glycosyl-flavonoids were discussed. The fourth chapter summarizes the research development on chemistry and pharmacology of medicine plants of B.chinensis and Iris species.

赤霉素、硫脲处理对黄精根状茎和须根发生的影响

为诱导无芽黄精根状茎产生不定芽,选择不同的化学物质进行诱导处理。结果表明,GA3处理效果明显,选用0.4 mg/L的GA3浸泡25 min对黄精根状茎不定芽的产生和须根生长有明显促进作用,高浓度抑制其生长;1%硫脲浸泡处理对黄精无顶芽根状茎萌发新芽和须根有明显的抑制作用。

Qualitative and semi-quantitative analysis of quaternary alkaloids in Coptis-scute herb couple by Electrospray mass Spectrometry

A practical solution of qualitatively analyzing quaternary alkaloids in coptis-scute herb couple by electrospray ionization mass spectrometry(ESI-MS) was developed. Without the complicated pretreatment of sample, the active ingredients including berberine, palmatine, coptisine, jatrorrhizine, epiberberine, and columbamine were identified and some relative content changing rules of alkaloids in coptis-scute couple were summarized in this article. The overall profiles of the complex extracts were obtained.

利用电喷雾质谱技术研究复方中药的配伍原则

Traditional Chinese Medicine (TCM) based on natural products is one important part of the Chinese civilization.Owing to the complexity of the composition,the study on medical effective components and curative effects are very difficult;Fuzi (Radix Aconiti Lateralis Praeparata) has been widely used for cardiotonic and analgesics in China,however,the component-aconitine in which is very toxic and may cause some side-effect.This paper reported the application of electrospray iohization mass spectrometric(ESIMS) technique on study of the compound prescriptions containing fuzi,Renshensini concoction and Baweidihuang concoction,to explore the mechanism of synergy between fuzi and other herbs.From chemical point of view,the detoxification mechanism can be attributed to the hydrolysis of the solubility of toxic diester-alkaloids from concoctions of fuzi.Radix Glycyrrhizae Praeparata,Rhizoma Zingiberis,Radix Ginsengp promote the hydrolysis to produce the less toxic monoester-alkaloids and Fructus Corni reduce the solubility of hypaconitine from fuzi.

A molecular marker that is specific to medicinal rhubarb based on chloroplast trnL/trnF sequences

"Da-Huang" (Radix et Rhizoma Rhei, medicinal rhubarb), a famous and important Traditional Chinese Medicine, has often been confused with the adulterant species in the same genus, Rheum. Through sequencing the trnL (UAA)/trnF (GAA) regions of chloroplast DNA of thirteen species of Rheum (three medicinal rhubarb species and ten adulterant ones), a molecular marker of the medicinal species was found. A pair of PCR primers based on the sequences, was thus designed, which amplified a highly specific DNA fragment in medicinal rhubarb exclusively, and absent in the adulterants at all under an optimized PCR condition.

Morfo-anatomia de espécies de Blastocaulon Ruhland (Eriocaulaceae)

Estudou-se a morfo-anatomia das raízes, caules, escapos e folhas de Blastocaulon albidum, B. prostratum e B. scirpeum, que ocorrem nos campos rupestres de Minas Gerais, Brasil. Verificou-se que as espécies apresentam raízes muito finas com epiderme unisseriada, duas-três camadas de células do parênquima cortical e tecidos vasculares com poucos elementos. Os caules em B. albidum e B. prostratum são aéreos, prostrados, ramificados, enquanto em B. scirpeum alguns indivíduos apresentam rizoma ou caules muito curtos. Constituem-se de epiderme, córtex e cilindro vascular. As folhas são delicadas, com células da epiderme com paredes finas, parênquima clorofiliano frouxo e reduzido número de feixes vasculares colaterais. Os escapos são cilíndricos, finos, não formam costelas e possuem apenas quatro feixes vasculares colaterais. As características observadas nessas espécies estudadas são típicas de plantas que ocorrem em locais úmidos e sombreados, ambientes onde é encontrada a maioria das espécies de Blastocaulon e ainda caracterizam anatomicamente o gênero, que se diferencia de Paepalanthus.

Prescriptions of Chinese medicinal herbs in Switzerland: the example of suan zao ren (Ziziphi Spinosae Semen)

Introduction According to the Swiss Health Survey 2007, 1.7% of the adult population use traditional Chinese medicine (including Chinese herbal medicine, but excluding acupuncture). In contrast to conventional drugs, that contain single chemically defined substances, prescriptions of Chinese herbs are mixtures of up to 40 ingredients (parts of plants, fungi, animal substances and minerals). Originally they were taken in the form of decoctions, but nowadays granules are more popular. Medium daily dosages of granules range between 8 to 12g. In a recent work we identified the most commonly used Chinese herbs (all ingredients are referred to as herbs for reasons of simplicity) and classical formulas (mixtures). Here we present a short overview and the example of suan zao ren (Ziziphi Spinosae Semen), which is used in the treatment of insomnia and anxiety and contains saponins that have been shown to increase sleep in animal studies. Material and Methods A random sample of 1,053 prescriptions was drawn from the database of Lian Chinaherb AG, Switzerland, and analysed according to the most frequently used individual herbs and classical formulas. Cluster analysis (Jaccard similarity coefficient, complete linkage method) was applied to identify common combinations of herbs. Results The most frequently used herbs were dang gui (Angelicae Sinensis Radix), fu ling (Poria), bai shao (Paeoniae Radix Alba), and gan cao (Glycyrrhizae Radix et Rhizoma); the most frequently used classical formulas were gui pi tang (Restore the Spleen Decoction) and xiao yao san (Rambling Powder). The average number of herbs per prescription was 12.0, and the average daily dosage of granules was 8.7g. 74.3% of the prescriptions were for female, 24.8% for male patients. Suan zao ren was present in 14.2% of all prescriptions. These prescriptions contained on average 13.7 herbs, and the daily dosage of granules was 8.9g. Suan zao ren was more frequently prescribed by practitioners of non-Asian than of Asian origin but equally often for female and male patients. Cluster analysis grouped suan zao ren with yuan zhi (Polygalae Radix), bai zi ren (Platycladi Semen), sheng di huang (Rehmanniae Radix) and dan shen (Salviae Miltiorrhizae Radix et Rhizoma). Discussion Prescriptions including suan zao ren contained on average slightly more herbs than other prescriptions. This might be due to the fact that two of the three most popular classical formulas with suan zao ren are composed of 13 and 12 herbs with the possibility of adding more ingredients when necessary. Cluster analysis resulted in the clustering of suan zao ren with other herbs of the classical formula tian wang bu xin dan (Emperor of Heaven’s Special Pill to Tonify the Heart), indicating the use of suan zao ren for the treatment of insomnia and irritability. Unfortunately, the diagnoses of the patients were unavailable and thus correlations between use of suan zao ren and diseases could not be analysed.

Effect of Arisaema erubescens (Wall) Schott rhizome extract on rheumatoid arthritis

Purpose: To investigate the anti-arthritic activity of the water extract of Rhizoma Arisaematis (WERA) using a collagen II -induced arthritis (CIA) rat model. Methods: CIA was induced in male Sprague-Dawley rats by intradermal injection of bovine collagen II in Complete Freund’s Adjuvant. The rats were treated with daily oral doses of WERA (100, 200, and 400 mg/kg) for 21 consecutive days. Methotrexate (MTX, 3 mg/kg), used as a positive control, was administered orally 2 times/week for 3 weeks. The severity of arthritis was evaluated using indices of paw swelling, arthritic score, body weight, thymus index, and spleen index. In addition, the serum levels of IL-1β, IL-6, IL-10, and TNF-α were measured. Results: All doses of WERA significantly inhibited paw edema (p < 0.01), decreased arthritis scores (p < 0.01) and spleen index (p < 0.05), and alleviated the weight loss associated with CIA in rats. Furthermore, TNF-α, IL-1β, and IL-6 serum levels were significantly decreased (p < 0.05) by all doses of WERA. By contrast, IL-10 serum levels were markedly increased (p < 0.05). Conclusion: WERA exerts therapeutic effects in CIA in rats by decreasing the serum levels of TNF-α, IL-1β, IL-6 and IL-10, suggesting WERA may be an effective candidate drug for treating human rheumatoid arthritis.