【作者】 张冬冬；

【导师】 陶文沂；

【作者基本信息】 江南大学 ， 微生物与生化药学， 2006， 硕士

【摘要】 自由基医学基础研究表明氧自由基引起的疾病已超过百种,近20年来,活性氧和自由基研究成为现代和生命科学的前沿和热点,评价和筛选具有强抗氧化活性的天然资源已成为生物学、医学和食品科学研究的新趋势。传统中药中存在着多种抗氧化成分,并在体内也有发挥抗氧化作用的可能性,所以引起了越来越多的研究关注。滨海县白首乌的主要来源为藦萝科植物耳叶牛皮消的块根。作为传统中药在江苏民间应用已有几百年历史。据报道白首乌有很多显著的药理作用,如抗癌、增强免疫、抗氧化的作用。本文以滨海白首乌为研究对象,研究了滨海白首乌抗氧化成分提取方法,分离与体内抗氧化效果的评价。本论文首先通过单因素实验结合响应曲面法对白首乌中抗氧化成分的提取工艺进行优化研究,确定了提取工艺的最佳条件为温度为:70.1℃,乙醇浓度为:75%,提取时间为:1.78 h,提取次数为两次。此条件下提取液的总抗氧化能力理论值为158.68U/g.用烘箱加速氧化法,测定油脂的POV值研究了探讨了此条件下的白首乌醇提液对大豆油、花生油、猪油氧化的抑制效果,并研究了醇提取液与Vc、柠檬酸的协同抗油脂氧化作用,结果显示乙醇提取液对三种油脂的脂质过氧化均有一定的抑制作用,和Vc有协同作用。通过静态和动态吸附性能比较,筛选出对白首乌抗氧化活性物质吸附性能最好的大孔树脂为HPD-400,最佳解吸附乙醇的浓度为80%。经HPD-400吸附纯化后,白首乌抗氧化提取物中总多酚含量提高了10.18倍,总黄酮提高了2.53倍。初步化学定性实验表明提取物中含有酚类、甾体苷类、黄酮类。用比色法检测了大孔树脂洗脱液清除羟基自由基、超氧阴离子自由基、DPPH的能力,抑制亚油酸的氧化能力。结果表明树脂洗脱物对羟基自由基、烷基自由基、DPPH自由基都有一定清除作用,但对超氧阴离子自由基无清除作用。用D-半乳糖建造小鼠衰老模型,给予不同浓度的树脂洗脱液,考察了实验小鼠脑指数、心指数、肝指数、脾指数、胸腺指数、肾指数变化,测定小鼠血清、脑组织和心组织中MDA、SOD的含量,测定小鼠肝组织中MDA、SOD、GSH-Px的含量。实验结果表明滨海白首乌大孔树脂洗脱物:(1)对小鼠脾指数有显著影响;(2)可提高衰老小鼠心组织SOD活力降低MDA水平(3)降低衰老小鼠脑组织MDA水平((4)提高肝组织SOD活力(P<0.05),降低MDA水平,可增强衰老小鼠肝GSH-Px活力,提高GSH-Px/LPO的比值(5)提高血清中SOD活力(P<0.05),降低MDA水平。更多还原

【Abstract】 Fundamental study in free radical iatrology showed that oxygen radicals relate to hundreds of diseases. In the last 20years, active oxygen and free radical have become the hot topics of modern life science, and selection of natural resources with strong antioxidative activities have become the new trends of biology, medical science and food science and technology. Antioxidants in Chinese traditional medicine may be useful in vivo and can be used to prevent oxidation impair. So more and more researchs focus on it.The root tuber of the Cynanchum auriculatum Royle ex Wight of the genus of Cynanchum is the main origin of the Baishouwu in Binhai county. As a traditional Chinese medicine, Baishouwu has been used in Jiangsu province for hundreds years.It has been reported that Baishouwu exhibited several potent pharmacological activities, including anti-tumor, antioxidant ability and immunologic enhancement. In this paper, optimum extract conditions of antioxidant from Baishouwu in Binhai county and purifucation conditions of absorbent resin column chromatography were studied, and the antioxidant activity were evaluated using vivo test.: Based on the results of one-factor-at-a-time-technique, a three-level Box-Behnken factorial design combining with response surface methodology(RSM) was employed to optimize the extraction condition of Baishouwu. When with two-time extraction, the extraction conditions of temperature, time and concentration of ethanol were set at 70.1℃,1.78 h and 75%,respectively. The activities of oxidation resistance of the ethanol extracts on the extraction condition of optimization were measured by lard, soybean oil and peanut oil as substrates. POV were measured by Oven Storage Method, and the synergistic actions were also investigated with viamin C and citric acid. The result showed that the optimum ethanol extracts have antioxidative activities to resist lipin-oxidation. Viamin C had synergistic antioxidative activities with ethanol extracts, but the synergistic effect of citric acid were not very significant.By comparing static and dynamic absorption performance, macroreticular resin HPD400 that had highest performance in absorbing antioxidative extract of Baishouwu and optimum absorption relieving agent 80% ethanol solution were selected. After absorption and purification by HPD400, the amount of total phenolic and total flavones compounds were increased by 10.18, 2.53 times. phenolic compounds, steroidal glycoside, and flavones in extract were confirmed by chemical qualitative analysis. Antioxidant activities of ethanol extract of macroreticular resin of Baishouwu(EEOMR) were evaluated using different in vitro tests: the DPPH radical scavenging activity, the reducing power, the inhibitory effect on hydroxyl radical and superoxide anion radical, the inhibitory effect on oxidation of linoleic acid. EEOMR was found to have antioxidant activities in all the model systems tested except the model of superoxide anion radical .The senior modle of Kunming-mice was built with D-galactose. Different doses of EEOMR were administered to the experimental mice. brain index, heart index, liver index, spleen index, kidney index and thymus index were measured. MDA and SOD of mice serum, brain, heart, liver and GSH-Px of mice liver, were measured. The results indicated that EEOMR could (1) effect on spleen index of mice (P<0.05), (2) increase the activities of SOD(P<0.01), reduce the level of MDA in heart, (3) reduce the level of MDA in brain (4) in crease the activities of SOD(P<0.05), GSH-Px/LPO ratio(P<0.01) and reduce the level of MDA in liver, (5) increase the activities of SOD(P<0.05), reduce the lever of MDA in serum.更多还原