【作者】 林秋凤；

【导师】 岑颖洲；

【作者基本信息】 暨南大学 ， 分析化学， 2004， 硕士

【摘要】 本文首次系统地对采自广东韶关的瑶药小紫金牛（Ardisia chinensis Benth）的化学成分和部分生物活性进行了研究。利用现代分离纯化技术和波谱鉴定手段对小紫金牛的化学成分进行了初步研究，并对小紫金牛水提物、醇提物和部分粗分离样品的抑菌、护肝和抗病毒活性进行了初步的探讨。 采用柱层析等现代分离纯化技术分析了瑶药小紫金牛（Ardisia chinensis Benth）的化学成分，并通过IR、¹HNMR、¹³CNMR、MS等结构鉴定手段从小紫金牛（Ardisia chinensis Benth）中共分离鉴定出7个化合物，它们的结构分别是（Ⅰ）豆甾-5，22-二烯-3β-醇、（Ⅱ）豆甾醇-3-β-D-吡喃葡萄糖苷、（Ⅲ）α-香树脂醇、（Ⅳ）羊毛甾-7，9（11），24-三烯-3β-醇、（Ⅴ）β-香树脂醇、（Ⅵ）羊毛甾-9（11），24-二烯-3β-醇、（Ⅶ）羊毛甾-8，24-二烯-3β-醇。以上化合物均为首次从小紫金牛中分离得到，并且除化合物豆甾-5，22-二烯-3β-醇和香树脂醇外，其它化合物均为首次从紫金牛属植物中得到。 采用水蒸汽蒸馏法从小紫金牛（Ardisia chinensis Benth）中提取其挥发性成分，再通过气相色谱-质谱联用（GC-MS）技术对其化学成分进行分离鉴定，并用气相色谱面积归一化法测定了各成分的相对含量。共鉴定了其中的50种化学成分，已成功鉴定的化合物占样品总量的90.6％，其主要成分是烯烃、酮、醇、酯、酸类物质。 采用微量稀释法测定了小紫金牛五种不同溶媒提取物及粗分离得到的七个样品对表皮葡萄球菌等6种细菌的最低抑菌浓度（MIC），探讨小紫金牛的体外抑菌活性。结果四种溶媒提取物及两种正丁醇分离样品均显示一定的抑菌效果，其中水萃取物对表皮葡萄球菌的抑菌效果最为显著，最低抑菌浓度MIC为0.313mg／ml，醇提物、乙酸乙酯萃取物、正丁醇萃取物及正丁醇的两个粗分离样品对表皮葡萄球菌显示出较好的抑菌活性，其最低抑菌浓度为0.625mg／ml。 采用CCl₄致小鼠实验型肝损伤为模型，探讨小紫金牛水提物对小鼠急性肝损伤的保护作用。结果与CCl₄模型组相比，小紫金牛水提物对CCl₄致小鼠血清谷丙转氨酶（ALT）、谷草转氨酶（AST）活性的升高有明显的降低作用（P＜0.01），其降酶效果与实验所用阳性药相当，表明小紫金牛水提物对CCI;致小鼠实验型肝损伤有明显的保护作用。但其不能明显降低肝组织MDA含量，表明其作用机制并非直接对抗脂质过氧化，而可能是作用于细胞膜，保护其不受氧自由基的破坏。 采用鸭乙型肝炎病毒（DHBV）体外原代鸭肝细胞（PDH）培养模型，探讨瑶药小紫金牛抗乙型肝炎病毒（HBV）的作用。应用胶原酶灌流DHBV感染雏鸭肝脏，制备原代培养的鸭肝细胞，斑点杂交法检测DHBV DNA。结果小紫金牛水提物对DHBV DNA表现出较强的抑制作用，IC50为273.5林g/ml。 上述活性研究证实瑶药小紫金牛有抑菌、降酶护肝、抗乙型肝炎病毒的作用，为进一步寻找与其功能主治相关的活性成分提供了科学依据。更多还原

【Abstract】 Yao Medicine Ardisia chinensis Benth, which occur in Shaoguan, Guangdong province, have been studied for the first time in this study. The constituents of Ardisia chinensis Benth have been studied by modern scientific technology. Biological activities, such as antibacterial, antivirus and acute liver injury, of aqueous extracts of Ardisia chinensis Benth and other samples that separated from it have also been studied.Seven compounds were separated from Ardisia chinensis Benth. Their structures were established on the basis of IR, 1HNMR, 13CNMR, and MS spectra to be stigmasta-5, 22-dien-3-ol (I), stigmasta-3--D-glucopyranosyl (II), Amyrin (III), lanosta-7, 9(11), 24-trien-3p-ol (IV), Amyrin(V), lanosta-9(11), 24-dien-3p-ol (VI), lanosta-8, 24-dien-3p-ol (VII). All these compounds were first separated from Ardisia chinensis Benth, and stigmasta-3-p-D-glucopyranosyl (II), lanosta-7, 9(11), 24-trien--3p-ol (IV), lanosta-9(11), 24-dien-3p-ol (VI), lanosta-8, 24-dien-3p-ol (VII) were first separated from Ardisia Sw.The volatile constituents of the essential oil of Ardisia chinensis Benth extracted by steam distillation have been studied by gas chromatography mass (GC-MS). A total of 50 compounds have been identified representing the 90.6% of the oil. The major constituents were found to be alkenes, ketones, alcohols, esters and acids.The antibacterial activities of different menstruum extracts of Ardisia chinensis Benth samples against 6 strains of bacteria such as Escherichia coli were investigated. Minimal inhibitory concentration (MIC) of 5 different menstruum extracts of Ardisia chinensis Benth and 7 samples that separated from Ardisia chinensis Benth to various bacteria was determined by the way of microdilution broth method in 96 wells plates. The results evaluates that the distilled water extracts shows a clear inhibitive effect to Staphylococcus epidermidids (MIC=0.313mg/ml), and the ethyl acetate extracts, the n-butanol extracts and two samples that separated from n-butanol extracts of Ardisiachinensis Benth shows some inhibitive effects to Staphylococcus epidermidids (MIC =0.625).The protective effects of the aqueous extracts of Ardisia chinensis Benth on mice acute liver injury induced by carbon tetrachloride (CCU) have been investigated. The results indicates that compared with those of the untreated group, the aqueous extracts of Ardisia chinensis Benth can significantly decreased the enzyme activities of alamine aminotransferase (ALT) and aspartate aminotransferase (AST) in liver injury in mice caused by carbon tetrachloride(CCl4), as the same effect with the positive control drugs. These enucleate that the aqueous extracts of Ardisia chinensis Benth shows a good protective effect on mice acute liver injury induced by carbon tetrachloride.The anti-HBV viral study of the aqueous extracts of Ardisia chinensis Benth was carried out in vitro by primary duck hepatocyte (PDH) culture system. The primary duck hepatocytes were obtained from DHBV-positive duck through collagenase perfusion, and the DHBV DNA was detected by dot-blot hybridization assay. The result evaluates that the aqueous extracts of Ardisia chinensis Benth shows a obvious effects on duck hepatitis B virus, IC50 is 273.5ng/ml.The above biological activitiy research substantiate that Ardisia chinensis Benth can work for bacterial, HBV virus and acute liver injur, and thus laid the foundations for studying the activity mechanism and relationship between the structure and the activity.更多还原