【作者】 崔堂兵；

【导师】 郭勇； 赵树进；

【作者基本信息】 华南理工大学 ， 发酵工程， 2002， 博士

【摘要】 银杏叶中的黄酮和萜内酯具有广泛的药理作用，银杏叶提取物（EGB）能治疗心脑血管疾病，增强记忆功能，预防和治疗老年痴呆，抑制过敏反应，抗氧化和清除自由基，增强人体免疫功能等，具有广泛的市场前景。因此，实现银杏细胞大规模培养生产银杏黄酮和萜内酯是保证今后市场需求的非常有前途的手段，而要实现银杏细胞大规模培养，提高细胞中银杏黄酮和萜内酯的产量是要解决的关键问题之一。本文在以前研究的基础上，对悬浮培养银杏细胞黄酮和萜内酯积累的调控、富含黄酮和萜内酯的银杏细胞提取物的制备以及药理活性等方面进行了研究。 1．L-苯丙氨酸和L-酪氨酸对细胞生长有无明显影响，添加适当浓度的L-苯丙氨酸和L-酪氨酸均能促进银杏黄酮的积累。1.0mM的乙酸钠能促进银杏细胞的生长，适当浓度的乙酸钠能促进银杏黄酮和内酯的积累。香叶醇对银杏细胞的生长无显著影响，适当浓度的香叶醇能促进内酯的积累。 2．添加水杨酸细胞生长影响不大，对银杏黄酮积累的影响促进作用不大，可提高银杏内酯的含量和产量。添加偏钒酸钠对银杏细胞的生长和银杏黄酮的积累起抑制作用，适当浓度的偏钒酸钠能促进银杏内酯的积累。添加茉莉酸甲酯抑制细胞的生长，适当浓度的茉莉酸甲酯可促进银杏黄酮和银杏内酯的积累。 3．当氯化氯胆碱添加浓度大于10mg.L^-1时，细胞生长受到抑制，10mg.L^-1的氯化氯胆碱对黄酮的积累有一定的促进作用，添加氯化氯胆碱能促进银杏内酯的积累。单萜抑制剂组合为松油醇10mg.L^-1+樟脑酸为5mg.L^-1时，银杏内酯产量达7138.73μg.L^-1，比对照提高了126.3％。 4．ATP对银杏细胞生长有促进作用，5’-AMP对细胞生长则起抑制作用；ATP对银杏黄酮和银杏内酯的积累有促进作用，5’-AMP则有抑制作用。苹果酸对银杏细胞生长的影响不大，适当浓度的苹果酸对银杏黄酮和银杏内酯的积累具有促进作用。 5．硝酸铈铵对银杏细胞的生长影响不大，高浓度则抑制细胞生长，10mg.L^-1的硝酸镧能促进细胞的生长。添加硝酸铈铵和硝酸镧对银杏黄酮积累的促进作用不明显，适当浓度的硝酸铈铵和硝酸镧能明显促进银杏内酯的积累。更多还原

【Abstract】 The flavones and terpenes（ginkgolides and bilobalide） in leaves of Ginkgo biloba L. have wide pharmacologic functions. The extract of ginkgo biloba（EGB） can cure cardiovascular and cerebrovascular diseases, enhance memory function, prevent and cure geriatric dementia, inhibit anaphylaxis, resist oxidation and clear free radical, enhance body s immunity function and so on. EGB has wide market outlook. In order to meet the need of pharmaceutical market for EGB, the large scale suspension culture of ginkgo biloba cells produces flavones and terpenes is a very prospective means, in which increasing flavones and terpenes output is one of key problem. In this article, on the basis of previous study, we studied the regulation of flavones and terpenes accumulation in ginkgo biloba cell suspension culture, the preparation the cell extract of ginkgo biloba with high content flavones and terpenes, the pharmacological activity. The results as follows.1. L-phenylalanine and L-tyrosine have no remarkable effects on cell growth, its s suitable concentrations can increase the accumulation of flavones. 1.0mM sodium acetate can enhance cell growth, the suitable concentrations of sodium acetate can increase flavones and terpenes accumulation. geraniol has no remarkable effect on cell growth, it s suitable concentrations can boost terpenes accumulation.2. Adding salicylic acid has no remarkable effects on cell growth and flavones accumulation, but can increase terpenes content and output. Sodium metavanadate inhibit cell growth and flavones accumulation, it’s suitable concentrations can promote terpenes accumulation, methyl jasmonate inhibit cell growth, it s suitable concentrations can increase flavones and terpenes.3. More than 10 mg.L^-1 chlorocholine chloride inhibit cell growth, 10 mg.L^-1 chlorocholine chloride can promote flavones accumulation, chlorocholine chloride can enhance terpenes accumulation. When the更多还原