【作者】 王宏伟；

【导师】 赵桂琴；

【作者基本信息】 承德医学院 ， 中药学， 2013， 硕士

【摘要】 乙型肝炎病毒(hepatitis B virus，HBV)感染可引起急性或慢性乙型肝炎，与肝硬化和肝癌密切相关，给人类的身体健康带来极大威胁。由于乙肝病毒的持续循环复制、病毒突变株的产生、机体免疫清除反应等原因，使得目前临床尚缺乏乙型肝炎的特效治疗药物。我国具有丰富的中草药资源，从中草药中寻找抗HBV活性好、高效低毒、新结构类型的天然化合物是开发抗乙肝天然活性药物的重要途径。素馨花为木犀科茉莉属植物素馨花（Jasminum officinale L. var.grandiflorum）的干燥花蕾，又名素馨针、耶悉茗花，世界各地广为栽培，具有疏肝解郁、行气止痛之功效。我国民间用其治疗消化不良、十二指肠球部溃疡及慢性肝炎、肝硬化等症。因此，通过相关文献调研，本文选取中医用于治疗肝病的中草药素馨花，对其化学成分进行分离纯化，并在细胞水平对其分离组分进行抗HBV活性筛选。首先，从素馨花70%乙醇提取物的氯仿、正丁醇萃取部位中分离纯化得到6个单体化合物，并通过1H-NMR、13C-NMR等波谱技术鉴定出其化学成分分别为：7-羟基-6-甲氧基香豆素（东莨菪内酯）、没食子酸乙酯、对苯二酚、2-(4-羟基-苄基)-苹果酸、红景天苷、对羟基苯乙醇，其中7-羟基-6-甲氧基香豆素（东莨菪内酯）、没食子酸乙酯、对苯二酚、2-(4-羟基-苄基)-苹果酸、红景天苷均为首次从素馨花中分离得到。进一步选择HepG22.2.15细胞为体外抗HBV活性评价模型，对上述素馨花70%乙醇提取物中的酚类化学成分进行体外抗HBV活性研究。结果显示：化合物没食子酸乙酯表现出体外抗HBV活性，对HepG22.2.15细胞分泌HBsAg具有一定的剂量依赖性抑制作用，IC50为62.5μg/ml。希望发现见诸报道三萜皂苷类和环烯醚萜苷类之外的具抗HBV活性的单体化合物，为中草药抗HBV活性成分研究作进一步探索，为素馨花药材的进一步开发利用提供科学依据。综上，本研究从素馨花中首次分离得到5个新的酚类化合物，并从中发现具有抗HBV活性的酚类化合物（没食子酸乙酯）。拓展了素馨花中抗HBV活性化学成分，即除了已知的三萜皂苷类和环烯醚萜苷类外，还有酚类组分，为素馨花抗HBV活性研究及成药开发提供了新的科学依据。进一步开展同为素馨花中分离得到的酚类化合物与三萜皂苷类、环烯醚萜苷类化合物的抗HBV活性比较性研究，将更全面的揭示素馨花的抗HBV活性物质基础及其构效关系；开展活性化合物衍生化研究，将进一步拓宽素馨花抗HBV活性药物研究思路；建立素馨花中的酚类化合物更优化的提取分离工艺和质量标准，并对其进行体内抗HBV活性评价，将更有效的发挥素馨花抗HBV活性的药用价值。更多还原

【Abstract】 Hepatitis B virus (HBV) infection can cause acute or chronic hepatitis B,which may lead to cirrhosis and hepatocellular carcinoma (HCC). Up to date,there are still no specific drugs to treat hepatitis B due to the continuous cycleof the replication of Hepatitis B virus, mutant strains of the virus and thebody’s immune clearance reaction. China has the treasure house of Chineseherbal medicine. Screening anti-HBV natural compounds with highbioactivity，low toxicity，new structures from Traditional Chinese Medicine isimportant to perform anti-HBV natural active pharmaceutical research.Jasminum officinale L．var．grandiflorum MuXi secco jasmine plant forJ.officinale dry flower buds, alias element, and zilpah mingyuan warm-heartedneedle flowers. It was cultured around the world due to the medicalapplication with the effect of Shu-Gan-Jie-Yu and gas painkillers．In ourcountryside, it is used to treat maldigestion, duodenal ulcer, chronic hepatitisbulbar urethra, cirrhosis and other disorders. Therefore, based on the previousliteratures, we chose Jasminum officinale L.var. grandiflorum as the rawmaterial due to its good efficacy of treating liver disease. The differentchemicals will be isolated from it and the anti-HBV activity will be evaluatedin HepG2.2.15cells to screen the specific chemicals for anti-HBV.After using chloroform and n-butanol to extract70%ethanol extract fromthe flower buds of J．officinale, six phenol monomer compounds were isolatedand purified. The structures of these compounds were confirmed by1H-NMR and13C-NMR. They are scopoletin(1), ethylgallate(2),1,4-benzenediol(3),2-(4-hydroxybenzyl)-malic acid(4), salidroside(5) and p-Hydroxy phenylet--hanol(6). Compounds1~5were isolated from Jasminum officinale at thefirst time.Then the anti–HBV activity of ethylgallate was evaluated by thepreliminary Anti–HBV activitiy assay using HepG2.2.15cell line, whichdisplayed the certain inhibition of the secretion of HBsAg with62.5μg/ml ofIC50.In summary, we isolated5new active fractions of phenol monomercompounds from J. officinale dry flower buds and found one of thesecompounds has the anti-HBV activity (ethylgallate). It proved that the phenolcompound also has bioactivity besides the well-known triterpenoid saponinsand iridoid compounds from J.officinale, which will be helpful to extend theresearch for the Traditional Chinese Medicine of J. officinale and to providethe scientific data for the new formulations. Moreover, it will find the basis ofanti-HBV activity of J. officinale and the relationship between the structureand activity by means of parallel investigation of phenolic compounds,triterpenoid saponins and iridoid compounds from J.officinale. Furthermore, itwill broad the scope of anti-HBV drugs from J.officinale throughderivatization. Finally, the medicinal value for anti-HBV of J.officinale will beenhanced by building the optimized extraction and separation technology andquality standards, even evaluation of anti-HBV activity in vivo.更多还原