【作者】 涂秋云；

【导师】 周春山；

【作者基本信息】 中南大学 ， 应用化学， 2008， 博士

【摘要】 中药现代化研究已成为目前医药领域的热点研究课题，采用现代化手段从中药分离制备活性成分是中药现代化研究的重要内容。本论文立足于药效，采用微波-半仿生法提取、大孔吸附树脂、柱层析法等中药现代化技术，首次系统地研究了连翘木脂素类成分的分离制备工艺，得到了易于工业化的工艺路线和条件参数；通过体外实验和动物体内实验研究探索了连翘木脂素抗衰老抗氧化、对脑损伤保护的生物学活性，取得了一系列有意义的成果。1．实验研究了连翘中木脂素类成分的不同提取工艺。采用传统的热浸提工艺提取连翘木脂素类成分，确定了提取工艺的优化参数。热浸提工艺提取过程平稳，设备要求不高，适合目前多数中药制药厂使用。研究了优化条件下的热浸提取动力学，结果显示提取过程是内扩散控制过程，提取表观活化能为19．74 kJ／mol，提取动力学方程为：采用微波-半仿生法模拟人体内消化道环境提取连翘木脂素类成分，实验显示证明微波-半仿生法提取效率高，优化条件下20分钟内通过两次提取，连翘木脂素的提取率高达87．5％，能耗低，前景广阔，产品的生物利用率得到提高。2．对连翘木脂素类成分进行了纯化研究。采用溶剂萃取-大孔吸附树脂-柱层析工艺纯化制备得到了纯度高达93．5％的精制连翘木脂素产品。采用大孔吸附树脂纯化连翘木脂素类成分，筛选出X-5树脂用于纯化，按照恒温25℃，上柱溶液木脂素类成分含量约为1000mg／L，吸附流速2BV／h，共吸附8 BV的上柱溶液；解吸时先用水淋洗，再用80％乙醇解吸，控制脱附流速为1BV／h，脱附剂的用量为3BV／批的优化工艺可以制备得到纯度达到82．5％的连翘木脂素产品，该纯化工艺过程简便，溶剂使用量少，对环境污染少，降低了成本，适合工业化扩大生产。通过对X-5树脂吸附行为的研究，确定X-5树脂吸附连翘木脂素类化合物过程主要是单分子的物理吸附，是放热的自发吸附过程，低温有利于树脂的吸附。Langmuir吸附等温方程能够很好地模拟X-5树脂等温吸附过程。3．研究了连翘提取物的抗氧化作用。通过研究得到连翘提取物的抗氧化活性强于化学合成氧化剂丁基羟基茴香醚，连翘提取物可以作为新型抗氧化剂加以开发。首次研究了连翘木脂素抗衰老抗氧化活性，体外实验显示连翘木脂素类化合物能够有效清除各种致衰老的自由基和成分。4．研究了连翘木脂素的对脑损伤保护的生物学活性。采用脑缺血再灌模型，通过比较连翘木脂素治疗组、对照组、假手术组实验动物小鼠的神经行为，分析血浆中超氧化物歧化酶、丙二醛、降钙素基因相关肽等成分的含量，观察小鼠海马CA1区中ET3和GFAP阳性细胞，证实连翘木脂素能减轻脂质过氧化反应、降低脑缺血再灌注海马区内皮素(ET)的表达、减缓蛋白质变性失活和膜屏障功能受损、减轻脑缺血再灌所致的神经损伤和脑神经元细胞死亡程度，具有脑缺血保护的生物学活性。研究结果提示连翘木脂素能够清除脑缺血损伤时产生的大量自由基是其具有此生物学活性的原因。更多还原

【Abstract】 Chinese medicine modernization research has been a hot subject in medicine field, while it is an important content that separating active components from Chinese medicine by modern technique. According to the pharmacodynamic activities, using the modern techniques of Microwave assisted-semi bionic extraction, macro porous resin seperation and column chromatography seperation, systematically study the sepertion art of lignan components, and getting an industrial art and related parameter;exploring the biological activities of lignan components on Anti-aging, oxidation resistance and protection of brain injuryby in vitro and in vivo experiments ,and get a series meaningful achievements.1. Studies of different extraction arts of lignan components of Fructus Forsythiae.Extracting lignan components of Fructus Forsythiae by traditional Heat Reflux extraction, definite optimize parameter of extraction art. The process of Heat Reflux extraction is stable, and needs common apparatus, the condition is adapt to most of Chinese medicine manufactures at present. Study dynamics of Heat Reflux extraction under the optimize condition, the result shows that extraction process is an internal spread controlling one, and the extraction apparent activation energy is 19.74 kJ/mol, extraction dynamics equation as following:Extracting lignan components of Fructus Forsythiae by microwave assisted-semi bionic extraction which imitated gastrointerstine surrounding in human, the results show that the extraction efficiency of microwave assisted-semi bionic extraction is higher, extration yield of lignans reach 87.5% under the optimic condition within 20 min and twice extraction, which means lower energy and higher yield, and has a good future.2. Study on purification of lignan components of Fructus Forsythiae. Preparing 93.5% lignans of Fructus Forsythiae by solvent extraction-macro porous resin seperation-column chromatography seperation.Selecting X-5 macroporous resin to purifying lignans of Fructus Forsythiae. The conditions are as following, temperature is 25 ======, content of lignans in solvent for column is 1000mg/L, absorbing velocity is 2BV/h, total volumn for absorbing is 8 BV;desorption process are as follow, water rinse at first, then by 80% ethanol and controlling the velocity for 1BV/h, getting 82.5% lignans after using 3BV ethanol under the optimize condition. This art is simple, using less solvent, causing less pollution , reducing cost, being adapt to industralization.According to study for the function of X-5 macro porous resin, determining the absorbing process of X-5 macro porous resin is physical absorbtion of single molecular, and it is spontaneous absorbtion, and it is easy to absorb under lower temperature.It can be imitated by Langmuir absorption isothermy equation that X-5 macro porous resin absorbs lignans process.3. Studying the oxidation resistance of Fructus Forsythiae extraction, and find its activity is better than anisole synthesized by chemical way, therefore, Fructus Forsythiae extraction can be developed as a new antioxidant. Studying the anti-aging and anti-oxidation of Fructus Forsythiae extraction, the results show lignan components can get rit of all kinds of free radicals which cause aging.4. Studying the biological activity of lignan components on protection of brain injury. Comparing with neuro-function of experimental rats in lignan components treatment group, control group, sham group after brain ischemia/reperfusion, measuring the content of erythrocuprein (SOD) , malondialdehyde (MDA) and calcitonin gene-related peptide (CGRP) in plasma of mice, observing the ET-3 and GFAP positive cell in hippocampus, results show that lignan components can decrease lipid oxidation and expression of ET-3, and decrease protein degeneration and membrain injury, and protecting ischemia/reperfusion injury. So we can conclude that it is one of important pathological processes in ischemia/reperfusion mice models that free radical formation and lipid peroxidation occur. Lignan compounds in fructus forsythiae can protect neurons in ischemia/reperfusion mice models from injury, and its antioxidation mechanism is perhaps related with the expression of calcitonin gene-related peptide.更多还原