- 學位論文
生物與非生物性誘導子對銀杏固定化細胞培養生產銀杏內酯B之影響
Effects of Biotic and Abiotic Elicitors on the Production of Ginkgolide B by Immobilized Cell Cultures of Ginkgo biloba
摘要
銀杏 (Ginkgo biloba)，杏仁科藥用植物，於中國傳統中醫上用於治咳平喘，而在現代藥理學實驗中亦證明銀杏萃取物確含有許多具醫療功能的成分，尤其以銀杏內酯B (Ginkgolide B，GB)最受人矚目，其具有防止血栓形成、抗血小板凝結、抗老化與治療轉移性癌症功能，市場需求量極大，因此以銀杏愈傷組織進行懸浮培養量產GB已勢在必行，但銀杏懸浮培養普遍遭遇GB產量極低問題，而二次代謝物多為植物防禦機制下產物，因此透過加入生物 (Yeast extract、Chitosan)與非生物 (Salicylic acid、Methyl jasmonate)誘導子激活防禦機制，以刺激GB產量提升。本研究以代表防禦機制強度的H2O2濃度和細胞存活率作為評斷指標，分析不同誘導子對GB產量的影響。 根據實驗結果顯示，茉莉酸甲酯 (Methyl jasmonate，MJ)與水楊酸 (Salicylic acid，SA)實驗結果最佳，其防禦機制強度相較於對照組分別提升了4.1倍與3.9倍，胞外GB產量提升5.0倍與3.6倍，而誘導子雖然會抑制細胞活性，但仍可保持8成存活率，而酵母萃取物 (Yeast extract，YE)與殼聚醣 (Chitosan，CH)雖然亦提升了3.5倍與4.4倍的防禦機制強度，但其胞外GB產量僅各提升了3.1倍，且存活率方面，除YE可維持8成存活率外，CH存活率大幅滑落至7成。初步的實驗結果說明誘導子確能有效激活銀杏防禦機制，但誘導子會對細胞造成抑制，因此選擇對細胞傷害較為溫和且GB產量高的非生物性誘導子較為合適。
並列摘要
Ginkgo biloba is a medicinal plant of the Ginkgoaceae family often used in traditional Chinese medicine to soothe coughs and shortness of breath. Modern pharmacology experiments have also confirmed that G. biloba extracts possess numerous components with medicinal properties, with the compound Ginkgolide B (GB) receiving the most attention. This compound protects against the formation of blood clots or thrombus and platelet clots, and possesses anti-aging and therapeutic qualities regarding metastasis. Consequently, market demand for GB is significant, and mass production of GB using suspension cultures from G. biloba calluses is a necessity. However, G. biloba suspension cultures generally provide a minimal yield of GB, and secondary metabolites are mostly byproducts of the plant defense mechanisms. Consequently, organic (e.g., yeast extract and chitosan) and inorganic (e.g., salicylic acid and methyl jasmonate) elicitors are used to activate defense mechanisms, thereby increasing the production of GB. This study used H2O2 concentration (an indicator of the strength of the defense mechanisms) and cell survival rates as indicators to determine the effects of different elicitors on GB production. The experimental results showed that methyl jasmonate and salicylic acid produced the best results. Their defense mechanism strengths were 4.1 times and 3.9 times stronger than those of the control group, with extracellular GB production increasing 5.0 times and 3.6 times, respectively. Although elicitors inhibit cell viability, 80% of the cells nevertheless survived. Furthermore, although yeast extract and chitosan increase the defense mechanism strength by 3.5 times and 4.4 times, respectively, extracellular GB production only increased 3.1 times. In addition, 80% of the cells survived the application of yeast extract; however, this figure dropped significantly to 70% when chitosan was applied. The initial experimental results demonstrated that elicitors effectively activate defense mechanisms in G. biloba; however, because elicitors inhibit cells, selecting inorganic elicitors that damage cells less and yield greater GB is preferred and more appropriate.
參考文獻
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