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题名: 棉花花总黄酮物质基础及抗阿尔茨海默病相关生物活性研究
作者: 赵永昕
答辩日期: 2012-12-04
导师: 胡克林、阿吉艾克拜尔.艾萨有机化学
专业: 有机化学
授予单位: 中国科学院大学
授予地点: 北京
学位: 博士
关键词: 棉花花 ; 黄酮类化合物 ; 阿尔茨海默病 ; 抗氧化
摘要: 本论文针对维吾尔药材棉花花总黄酮有效部位及其黄酮类化合物物质基础进行了抗阿尔茨海默病相关生物活性及作用机理研究。1. 棉花花总黄酮有效部位中黄酮类化合物的分离及结构鉴定棉花花总黄酮的制备来自于实验室成熟稳定的工艺获得,采用聚酰胺柱色谱及
LH20 凝胶色谱柱结合的方式共分离含量较高的8 个黄酮类化合物,经核磁谱图分析确定8 个化合物的化学结构。2. 棉花花总黄酮及黄酮类化合物体外抗氧化作用研究采用DPPH 和ABTS 两种自由基清除法来研究棉花花总黄酮及其黄酮类化合物体外清除自由基活性能力,并进行了脂质过氧化的研究,实验结果显示棉花花总黄酮及其黄酮类化合物均具有明显的抗氧化作用,并针对实验结果对黄酮类化合物的结构进行构效关系分析。3. 棉花花总黄酮及黄酮类化合物对乙酰胆碱酯酶抑制活性研究乙酰胆碱酯酶抑制剂作为阿尔茨海默症(AD)的治疗药物,仍然是药物化学的研究热点之一,有研究表明黄酮类化合物也具有乙酰胆碱酯酶抑制的活性。本研究采用体外方法对棉花花总黄酮及黄酮类化合物对乙酰胆碱酯酶抑制活性进行了研究,同时采用AUTODOCK 软件对小分子黄酮类化合物配体和大分子乙酰胆碱酯酶进行了分子对接研究,并给出结合能大小。4. 棉花花总黄酮及黄酮类化合物对tBHP诱导PC12细胞氧化应激的保护作用活性氧被认为是很多疾病致病机理的重要因素,有研究表明氧化应激诱导细胞损伤可能是AD神经退行性病变的重要原因,本研究以PC12细胞为研究对象,采用叔丁基过氧化氢(tBHP)诱导PC12细胞损伤建立AD细胞模型,MTT法检测棉花花总黄酮及黄酮类化合物对PC12细胞的损伤保护作用。5. 棉花花总黄酮及黄酮类化合物对H2O2诱导的人成神经细胞瘤SK-N-BE-2细胞损伤及作用机理研究Bcl-2为凋亡抑制因子,定位于线粒体,在细胞内凋亡信号转导中起关键作用,本论文建立H2O2诱导的SK-N-BE-2凋亡模型,研究棉花花总黄酮及黄酮类化合物对SK-N-BE-2的损伤保护作用情况,并研究SK-N-BE-2受H2O2诱导后其体内抗凋亡蛋白Bcl-2基因水平的变化。6.异槲皮素标准品研究天然产物标准样品在天然产物的质量控制中,是确认产品质量必不可少的标准参照物。异槲皮素为棉花花总黄酮中分离得到的一个化合物,具有清除自由基、抗氧化作用,目前国内外没有市售的异槲皮素标准样品,也未见分离制备异槲皮素对照品的文献报道,我们对其进行标准样品的研究,可用它作为标准参照物对天然产物质量进行评价,保证分析测试质量,使测试结果具有准确性与合法性。
英文摘要: In this paper,we investigated the material basis of total flavonoids from Flos Gossypii. and their activity of anti-Alzheimer's disease. 1. The separation and structural identification of flavonoids from Flos Gossypii. Total flavonoids of Flos Gossypii were from mature and stable process route which were done by our lab study .Eight flavonoids were separated by polyamide and sephadex LH-20 column chromatograph. The structural identifications of compounds were carried out using 1HNMR, 13CNMR and high-performance liquid chromatography multistage tandem mass spectrometry as well as comparing with our previous report. 2. Research on antioxidant activity of total flavonoids and flavonoids from Flos Gossypii. The antioxidant activities of compounds as well as total flavonoids from Flos Gossypii were examined by two tests: DPPH and ABTS radical activity. The data showed that eight compounds and total flavonoids had the antioxidant activity. We also discussed the structure-activity relationship of these compounds. 3. Research on inhibition of acetylcholinesterase (AchE) of total flavonoids and flavonoids of Flos Gossypii. AchE inhibitors were therapeutic drug of treating AD, which were still research hotspot of chemical chemistry. Some study showed that flavonoids have AchE inhibition activity. In this paper we investigated the AchE inhibition of total flavonoids and then flavonoids from Flos Gossypii and analyzed the molecular docking energy between ligands (flavonoids) and receptor (AchE). 4. Research on protection action of tBHP- induced PC12 cell injure from total flavonoids and flavonoids of Flos Gossypii. Reactive oxygen species are believed to play a very significant role in the pathogenesis of several diseases including AD. There are some studies suggesting that oxidative stress is one of the reason of AD degenerative change. In this paper, total flavonoids and flavonoids of Flos Gossypii were investigated the protection of PC12 cell injure induced by tert-Butyl hydroperoxide (tBHP). 5. Research on protection action and mechanism of H2O2-induced SK-N-BE-2 cell injure and mechanism of action from total flavonoids and flavonoids of Flos Gossypii. Bcl-2, which is a kind of apoptosis inhibiting factor, was located in mitochondria and it played pivotal role in intracellular apoptosis signal. In this paper we researched on the protection action of SK-N-BE-2 cell apoptosis from total flavonoids and flavonoids of Flos Gossypii and investigated the Bcl-2 gene expression change. 6. Research on isoquercetin standard substance Natural product standard substance is very important in verifing the quality of product during the quality control of natural product. Isoquercetin is one of component of total flavonoids from Flos Gossypii. , which has the antioxidant activity and eliminate the free radical .In present, isoquercetin has not reported in the market at home and abroad. Understanding it through research, we can take it as the standard substance to evaluate and make sure the analytical result of the quality of product accurate and legal.
内容类型: 学位论文
URI标识: http://ir.xjipc.cas.cn/handle/365002/2466
Appears in Collections:资源化学研究室_学位论文

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作者单位: 中国科学院新疆理化技术研究所

Recommended Citation:
赵永昕. 棉花花总黄酮物质基础及抗阿尔茨海默病相关生物活性研究[D]. 北京. 中国科学院大学. 2012.
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