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耳叶牛皮消化学成分和活性研究

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Ⅰ.Genus Cynanchum: A Review of Chemical Constituents and Pharmacological Activities

1.1 Introduction

1.2 Chemical Constituents

1.2.1 Steroids

1.2.2 Alkaloids

1.2.3 Acetophenones

1.2.4 Triterpenoids

1.2.5 Flavonoids

1.2.6 Others Constituents

1.3 Pharmacological Activities

1.3.1 Anti-tumor Activity

1.3.2 Immuno-modulatory Activity

1.3.3 Anti-epileptic Activity

1.3.4 Anti-virus Activity

1.3.5 Antioxidative Activity

1.3.6 Other Activities

1.4 Conclusions

References of Chapter I

Ⅱ.Chemical Constituents of Cynanchum auriculatum

2.1 Introduction

2.2 Extraction, Isolation and Purification

2.3 Chemical Structures of Idetified Compounds

2.4 Structure Elucidation

2.4.1 New C21 Steroidal Glycosides

2.4.2 New Acetophenone

2.4.3 Known Compounds

2.5 Experimental Section

2.5.1 Plant Material

2.5.2 General Experimental Procedures

2.5.3 Acid Hydrolysis of Steroidal Glycosides

2.5.4 GC Analysis for Determination of Glucose Absolute Configuration

2.5.5 Physico-chemical Property and NMR Data

2.6 Discussion

2.6.1 The Characteristics of C21 Steroidal Glycosides from Cynanchum auriculatum

2.6.2 The Isolation and Purification Flowchart of C21 Steroidal Glycosides

References for Chapter Ⅱ

Ⅲ.Bioactivies of Cynanchum auriculatum

3.1 Introduction

3.1.1 The Traditional Use of Cynanchum auriculatum

3.1.2 The Strategy of Bioactivity Studies on Cynanchum auriculatum

3.2 Anti-tumor Constituents

3.2.1 Anti-tumor Activity of Extract and Fractions

3.2.2 Anti-tumor Activity of Steroidal Glycosides

3.2.3 Qualitative Structure-activity Relationship of Steroidal Glycosides'Anti-tumor Activity

3.2.4 Experimental Section

3.3 Neuroprotective Constituents

3.3.1 Neuroprotective Effect against Glutamate Excitotoxicity

3.3.2 Anti-Alzheimer's Aβ42-induced Toxicity in Drosophila melanogaster

3.3.3 Mechanism Investigation of Neuroprotective Constituent-Cynandione A

3.4 Gastroprotective Constituents

3.4.1 Introduction

3.4.2 Gastroprotective Activity of Extract and Fractions

3.4.3 Gastroprotective Activity of Acetophenones from Cynanchum auriculatum

3.5 Discussion

3.5.1 Anti-tumor Constituents

3.5.2 Neuroprotection Constituents

3.5.3 Gastroprotection Constituents

References for Chapter Ⅲ

Papers and Patents

Acknowledgements

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摘要

本论文研究了萝藦科鹅绒藤属植物耳叶牛皮消(Cynanchum auriculatum Royle exWight)的化学成分和生物活性,着重研究了其中的C21甾体皂苷和苯乙酮类成分,以及抗肿瘤、神经保护和胃保护活性。应用常压柱色谱、中压和高压制备液相等分离技术从耳叶牛皮消乙醇提取物中分离纯化了90个化合物,并运用UV,IR,ESI-MS,1D-NMR,2D-NMR等光谱技术和物理、化学方法鉴定了其化学结构,其中19个为新化合物(18个新C21甾体皂苷,1个新苯乙酮),61个化合物为首次从该植物中分得。本文给出了这90个化合物的结构。 通过抗肿瘤、神经保护和胃保护三方面的药理研究,发现耳叶牛皮消总提物和其氯仿部位即总皂苷在体外人体肿瘤细胞株细胞毒实验和荷瘤小鼠体内肿瘤生长抑制实验中都显示了显著的抗肿瘤活性,因此确证了耳叶牛皮消的抗肿瘤活性,并发现总甾苷是其抗肿瘤活性部位。进一步的单体活性筛选发现甾体苷元caudatin及其糖苷具有显著的抗肿瘤活性,构效关系研究发现12位异戊酸基团对C21甾体皂苷的细胞毒活性具有较大贡献,而3位去氧糖糖链和12位肉桂酸基团则会降低C21甾体皂苷的细胞毒活性。在体外培养的神经元细胞和PC12细胞上进行的神经保护实验,发现苯乙酮化合物cynandione A具有显著的抗L-谷氨酸诱导神经兴奋性毒性作用和抗H2O2诱导的神经损伤作用,其作用机制可能与上调ERK基因表达从而促进细胞存活相关;同时cynandione A还在Aβ高表达的转基因果蝇模型上显示了较好的改善果蝇受损爬行能力活性,在早老性痴呆治疗领域显示了较好的临床应用前景。同时还发现高剂量的cynandione B和cynantetrone具有显著的、不依赖谷氨酸的神经损伤作用。在胃保护实验中,采用吲哚美辛、乙醇引起的大鼠胃粘膜急性损伤和组胺引起的大鼠胃酸分泌增加模型,发现耳叶牛皮消氯仿部位即总甾苷具有显著的胃保护活性,特别是发现其抑制胃酸分泌活性非常显著,作用强度与阳性药西咪替丁相当。

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