臺灣博碩士論文加值系統

銀杏是中國最古老的藥用植物，其銀杏葉中所包含的銀杏內酯(ginkgolide A、B、C、J、M、簡稱為GA、GB、GC、GJ、GM），皆具有拮抗血小板活化因子（Platelet-activating factor，PAF）的作用以及保護神經細胞的功能，其中以GB的效力最強，目前被認為是最有臨床應用前景的天然PAF受體拮抗劑，對於血栓、心腦血管疾病、心律失常、平喘、支氣管炎、老年癡呆症、過敏反應等預防和治療具有一定作用。故其被廣泛地應用於醫藥、健康食品等工業。本研究乃嘗試利用植物組織培養技術建立銀杏癒合組織培養系統，並測定其所含成份ginkgolide B。再藉由懸浮細胞體系大量生產銀杏細胞及其二次代謝產物。其結果顯示，在銀杏癒合組織的誘導培養基上，以添加2.0mg/l NAA、0.5mg/l kinetin及 3% sucrose之MS 基礎鹽類培養基之誘導率最高。而在癒合組織的繼代培養上則以使用1/2MS基礎培養基鹽類添加3% sucrose、2mg/ l NAA、0.5mg/l kinetin及0.75% agar之培養基（pH5.7±0.1），於黑暗及25±1℃之恆溫下培養最佳。此外在液態懸浮細胞培養方面，使用添加2mg/l NAA、 0.5mg/l kinetin及3%sucrcose之液體1/2MS培養基（pH5.7±0.1），接種3ml PCV/125ml medium之細胞初體積，於100rpm轉速下進行培養其細胞生長最為良好。此外在銀杏二次代謝產物ginkgolide B的含量分析上，由莖誘導所得癒合組織之GB含量最高。

Ginkgo biloba is an ancient Chinese medicinal plant. The Ginkgo Biloba leaves contain mainly ginkgolide（ginkgolide A, B, C, J , and M, abbreviated as GA, GB, GC, GJ, and GM）are selective platelet-activating factor（PAF）antagonist and posses the neuro-protective effect. Especially, GB has the most strong potent among these terpenoid and is now used as the antagonist for the prevention and treatment of thrombus , illness of blood vessel of heart and brain arrhythmia , asthma ,bronchitis , senile dementia and allergic reaction, and is also widely used in pharmaceutical, health food industry and others. The objective of the present investigation was to establish an efficient ginkgo biloba callus and to analyze the content of ginkgolide B. The suspension cell culture was established to produce ginkgo biloba cell and secondary metabolite in large scale. The results obtained are summarized as follows. The MS basal medium supplemented with 2.0mg/l NAA 、0.5mg/l kinetin and 3% sucrose was found to yield the highest induction rate of callus. The 1/2MS basal medium supplemented with 3%sucrose 、2mg/l NAA 、0.5mg/l kinetin and 0.75% agar （pH5.7±0.1）incubated of culture in dark at 25±1℃ was found to be optimum for the proliferation of callus. The 1/2 MS basal medium supplemented with 2mg/l NAA 、0.5mg/l kinetin and 3% sucrose （pH5.7±0.1） inoculated with 3ml PCV/125ml initial cells shook with 100rpm was found to proceed cell suspension culture significantly. Moreover，the callus induced from the stem was found to produce the highest content of secondary metabolite ginkgolide B.

中文摘要………………………………………………………………….I
英文摘要………………………………………………………………..III
總目錄…………………………………………………………………...V
表目錄…………………………………………………………………VII
圖目錄………………………………………………………………...VIII
第一章 緒言………………………………………………….………….1
第二章 文獻回顧………………………………………………………..7
2.1銀杏概述………………………………………..………………7
2.1.1 銀杏………………………………………………………..7
2.1.2 銀杏葉的成份與藥理作用………………………………..7
2.1.3 銀杏之安全有效的服用法………………………………12
2.2植物的組織培養………………………………………....…….15
2.2.1 緒論……………………………………….……..……….15
2.2.2 植物細胞之構造與機能…………………………………15
2.2.3 植物細胞之分化全能性…………………………………16
2.2.4 植物組織培養技術………………………………………18
第三章 實驗部份…………………………………..…………………..37
3.1 材料和方法……………………...……………..…………..37
3.2 HPLC銀杏內酯之化學成份測定…………….……….…..40
第四章 結果與討論………………………………….………………...42
4.1 銀杏癒合組織培養之研究…………….….….…………....42
4.2 銀杏液體懸浮培養之研究…………….….…….………....47
4.3 二次代謝物Ginkgolide B之含量測定……………………55
第五章 結論……………………………………………………………69
參考文獻………………………………………………………………..71

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