【作者】 何春年；

【导师】 肖培根； 彭勇；

【作者基本信息】 中国协和医科大学 ， 生药学， 2010， 博士

【摘要】 芍药科(Paeoniaceae)系单属科,由芍药属(Paeonia)从毛茛科(Ranunculacea)独立而来。芍药属约35种,划分为3个组,即牡丹组、北美芍药组和芍药组,中国是芍药科植物的起源、演化、分化发展及多样性中心,特别是其中的牡丹组为我国特有植物。为了更深入地探讨芍药属植物的药用亲缘关系,阐明该属植物的分类学地位和种间关系,为资源利用和保护提供依据,我们首先在对牡丹种子进行系统的化学成分研究基础上,系统分析中国产芍药属主要药用植物种子的化学成分含量与分布特点,通过数据统计分析揭示其化学分类学意义；通过快速色谱分析白芍、赤芍和牡丹皮的指纹特征,采用化学计量学方法分析三味药材化学成分的差异,探讨三味药材药性的差异与化学成分差异之间的相关性；并详细对芍药属牡丹组和中国产芍药组植物的色谱指纹图谱进行研究和分析,运用统计分析方法进行化学分类学探讨,揭示芍药属植物根部主要次生代谢产物分布特点。在上述研究的基础上,探讨芍药属(科)的药用植物亲缘学关系,同时根据药用植物亲缘学理论对牡丹种子资源利用的前景进行展望。本文对牡丹(P.suffruticosa)种子进行了较系统的化学成分研究。采用硅胶柱色谱初步分离,结合反相色谱和Sephadex LH-20色谱等多种分离纯化手段,对牡丹成熟种子的乙醇提取物进行分离,通过多种波谱学技术鉴定了其中31个化学成分,其中芪类成分11个,分别为：Resveratrol (E)-form (1)、Resveratrol (Z)-form (2)、trans-ε-Viniferin (3)、cis-ε-Viniferin (4)、Suffruticosol A (5)、Suffruticosol B (6)、Suffruticosol C (7)、Gnetin H (8)、cis-Gnetin H (9)、trans-Suffruticosol D (10)和cis-Suffruticosol D (11);单萜苷类11个,分别为：6’-O-β-D-葡萄糖芍药内酯苷(18)、芍药苷(19)、氧化芍药苷(21)、β-gentiobiosylpaeoniflorin (22)、8-去苯甲酰芍药苷(23)、8-Debenzoylpaeonidanin (24)、Debenzoyl albiflorin (26)、Pyridylpaeoniflorin (27)、Piperitone-4-en-9-O-β-D-glucopyranoside (28)、1-O-β-D-Glucopyranosylpaeonisuffrone(29)和芍药内酯苷(31)；黄酮类4个,分别为：芹菜素(13)、木犀草素(14)、槲皮素(15)和山萘酚(16)；其他类5个,分别为：对羟基苯甲醛(12)、苯甲酸(17)、1-O-α-D-乙基甘露糖苷(20)、蔗糖(25)和1-O-β-D-对羟基苯甲酰葡萄糖苷(30)其中是9、10、11、22、27和28是新化合物,其余有3个为本属植物首次分离得到,有9个为本种植物首次分离得到。另外,对其中10种芪类化合物(1,3-11)的活性进行了初步研究,清除DPPH自由基和诱导Keap1-Nrf2-ARE信号通路均具有适度的活性,表明这些化合物可能会在抗癌、抗衰老、抗炎中发挥作用。在上述化学成分研究的基础上,首次采用HPLC-DAD技术分别建立了同时测定10种芪类(化合物1、3-11)和8种苷类(化合物18、19、21、22、27、28、30和31)化合物的含量测定方法,并对芍药属植物种子46份样品(其中牡丹组32份,6种4变种1杂种；芍药组14份,3种)进行了含量测定,利用主成分分析方法对测定结果进行进一步的数据分析,对芍药属的植物的化学分类学进行了探讨。首次对牡丹、凤丹和芍药种子中主要化学成分(化合物1、3-11、19和22)的季节性动态变化进行了研究,发现这些化合物在种子生育期内呈明显规律性,10种芪类化合物在种子近成熟时(8月)才大量合成,β-gentiobiosylpaeoniflorin (22)与芍药苷(19)有相互转化的可能。在对种子部位研究的基础上,我们进一步采用HPLC技术对传统药用部位根/根皮进行了指纹图谱研究。首先采用HPLC整体柱快速分析比较了白芍、赤芍和牡丹皮三味药材共81份样品的指纹图谱,利用指纹图谱数据处理软件比较他们的相似度,建立了色谱图谱共有模式,利用主成分分析方法进行数据分析,三味药材能够很好地区分,在指认了13种常见主要化学成分的基础上,比较了影响他们异同的物质基础。首次利用HPLC-DAD方法对中国产芍药属植物52份样品(其中牡丹组8种29份样品,芍药组共有6种23份样品)根部的主要次生代谢产物进行了全面分析,分别建立了牡丹组和芍药组植物的色谱图谱共有模式,利用主成分分析方法分别对两组植物进行了比较,两组植物均显示了不同程度的区分,指认了15种常见主要化学成分,比较影响他们异同的物质基础,对芍药属的化学分类学具有一定的意义。本文还对芍药属的文献资料进行了较全面的综述,包括化学成分、药理活性和分析方法等,并对芍药属植物的化学分类学和药用植物亲缘学进行了探讨。更多还原

【Abstract】 Paeonia, the only genus in the family of Paeoniaceae, comprises ca.35 species, including three sections:Moutan, Oneapia, and Paeonia. The section Moutan, with eight woody species, is confined to a small area in China.Kinds of chromatogramphic methods were employed to isolate and purify compounds from the seeds of Paeonia suffruticosa.31 compounds were isolated from the ethanol extract of Moutan seeds, these compounds were elucidated based on spectral analysis and comparison with spectroscopic data reported in the literatures, including 11 stilbenes,11 monoterpenoid glycosides,4 flavanoids and 5 other types compounds. Among these identified compounds,6 compounds of them were new compounds,9 were the first time to be isolated from this plant species, and 3 were the first time to be isolated from the genus Paeonia.The six new compounds were named as cis-gnetin H (9), trans-suffruticosol D (10), cis-suffruticosol D (11),β-gentiobiosylpaeoniflorin (22), pyridylpaeoniflorin(27), and piperitone-4-en-9-O-β-D-glucopyranoside (28). Structures of the known compounds were identified as resveratrol (E)-form (1), resveratrol (Z)-form (2),trans-ε-viniferin (3), cis-ε-viniferin (4), suffruticosol A (5), suffruticosol B (6), suffruticosol C (7), gnetin H (8), p-hydroxybenzaldehyde (12), apigenin (13), luteolin (14), quercetin (15), kaempferol (16), benzoic acid (17),6’-O-β-glucopyranosylalbiflorin (18), paeoniflorin (19), 1-O-a-D-ethyl-mannopyranoside (20), oxypaeoniflorin (21),8-debenzoylpaeoniflorin (23), 8-debenzoylpaeonidanin(24), sucrose (25), debenzoyl albiflorin(26),1-O-β-D-glucopy ranosylpaeonisuffrone (29), 1-O-β-D-(4-hydroxybenzoyl)glucose (30), and albiflorin (31).Compounds 1,3-11 were assayed for their DPPH free radical scavenge activities and inducibility of the Keapl-Nrf2-ARE signaling pathway, the results showed these compounds were possessed moderate activities, and may be produce a marked effect to anticancer, anti-aging, and anti-inflammatory.A facile method based on high performance liquid chromatography coupled with photodiode array detector (HPLC-DAD) was developed for the simultancous quantitative determination of ten stilbenes (1,3-11) and eight glycosides (18、19、21、22、27、28、30, and 31) in 46 batches of samples including Sect. Moutan with 32 batches and Sect. Paeonia 14 batches from 9 species. Principal component analysis (PCA) were applied to classify the Paeonia samples and chemotaxonomic characteristics were proposed.The content of major compounds (1,3-11,19, and 22) in the seeds of P.lactiflora, P. suffruticosa, and P.ostii collected from different ontogenetic stages were detected by HPLC-DAD. The results showed these compounds presented conspicuous regularity, stilbenes synthetized abundantly until seed approaching period of maturation, and compounds 19 and 22 can be reciprocal transformation.Compared the distinctness of HPLC-DAD fingerprints chromatography among Radix Paeonia Alba, Radix Paeonia Rubra and Cortex Moutan.81 batches samples were rapid analysis by HPLC equipping Chromolith column, the chemometrics procedures, including similarity analysis (SA) and PCA were applied to classify these three categories samples. Consistent results were obtained to show that these samples could be successfully grouped in accordance with their types. Furthermore,13 marker constituents were screened out to be the main chemical composition.Compared the distinctness of root/root bark chemical fingerprints chromatography among Sect. Moutan and Sect. Paeonia.52 batches samples including 29 Sect. Moutan from 8 species and 23 Sect. Paeonia from 6 species were multi-analysis, the chemometrics procedures, including similarity analysis (SA), PCA were applied to classify these Sect. samples. Consistent results were obtained to show that these samples could be temperately grouped in accordance with their species. Furthermore,15 marker constituents were screened out to be the main chemical composition.In addition, advance in the research of phytochemical, biological, and analytical method about quality control of medicinal plants from Paeonia genus were reviewed whith considerable references. Furthermore, chemotaxonomy and pharmacophylogeny of Paeonia genus was investigate.更多还原