藥用石斛對RAW264.7細胞產生一氧化氮的調節作用與組織培養之研究; Studies on the nitric oxide by RAW264.7 cell & tissue-culture of Dendrobii herba

中國醫藥大學機構典藏 China Medical University Repository, Taiwan > 藥學院 > 藥物化學研究所 > 博碩士論文 > Item 310903500/24394

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題名:	藥用石斛對RAW264.7細胞產生一氧化氮的調節作用與組織培養之研究;Studies on the nitric oxide by RAW264.7 cell & tissue-culture of Dendrobii herba
作者:	何惠雅;Ho Hui-Ya
貢獻者:	中國醫藥學院藥物化學研究所
關鍵詞:	藥用石斛;組織培養;一氧化氮;石斛屬;Dendrobii herba;tissue-culture;nitric oxide;Dendrobium
日期:	1992
上傳時間:	2009-12-23
摘要:	中文摘要藥用石斛最早收載於神農本經上品，歷代諸家本草皆有記載，主要功效為清熱生津，滋陰養胃，清肝明目等，其主要活性成分為生物鹼與多糖。本研究以各種藥用石斛提取物對LPS(Lipopolysaccharide)與IFN-γ(Inteferon-γ)所誘導巨噬細胞產生一氧化氮的影響，作為生物活性篩選指標，針對具有此種生物活性的石斛等物種，進行植物組織培養，其目的作為建立大量的栽培方法，並提供進一步篩檢生理活性成分的材料。結果如下： 1. 15種石斛氯仿萃取率中，以霍山石斛最高(6.63﹪)，其次為台灣產美花石斛(4.99﹪)與大陸產串珠石斛(4.82﹪)；水的萃取率以台灣產綠黃花石斛(27.72﹪)與霍山石斛最高(25.27﹪)。 2.氯仿萃出物對於LPS與IFN-γ誘導RAW264.7巨噬細胞產生一氧化氮的抑制率以日本石斛最高 (73.6﹪)；水萃出物對一氧化氮的產生促進作用最高者為鐵皮石斛 (-44.8﹪)。 3.日本產石斛的外殖體培養中，以B5的基本鹽類培養基，添加BA的側芽生長較好。 4.從4公分側芽的莖切取1公分長的莖段石斛的莖培養於含2.5 mg/L BA + 0.5 mg/L NAA之B5培養基，再生側芽數多且適合作為成苗培養的材料。 5.成苗培養：再生側芽(大於1公分)移植G-3培養基(3 g/L Hyponex 1＋15﹪馬鈴薯汁＋15﹪香蕉汁＋15 ﹪椰子汁＋0.5 mg/L BA＋3 g/L蔗糖＋2 g/L活性碳) 培養90天後可長成4.5公分高，根2~3條，根長5~6公分的苗體，供馴化出瓶之用。 6.類原球體培養以2.5 mg/L BA＋3﹪蔗糖的B5固體培養基，能產生較多類原球體。 7.馴化條件：瓶苗置於栽種溫室30天後，脫瓶移出的存活率最高 (85﹪)。 8.將石斛培養苗移出三角瓶外，栽種於不同比例之混合介質與容器中，發現種於蛇木屑：石塊＝1：1組合的介質，並以700 mL透明飲料瓶栽種的石斛，可適合平地溫室大面積種植。 9.以品種交配所得種子與原種的石斛種子，檢討其種子培養到栽種的條件，提供今後藥理篩選材料，探討最佳種植的品種。 10.三種栽種石斛(台灣綠黃花石斛，日本石斛，霍山石斛)的鮮重與乾重均以日本石斛最佳。; The stems of several Dendrobium species are used in traditional Chinese medicine as a Yin tonic to nourish the stomach, promote the production of body fluid and reduce fever. In the present study we have established bioassay tests on Dendrobii extracts for production of nitric oxide by murine macrophage-like cell line (RAW264.7) activated with LPS (Lipopolysaccaride) and INF-γ (Interferon-γ). We have developed protocols for in vitro propagation of Dendrobium plants and the results obtained were summarized as follows 1.Out of the fifteen species, Dendrobium hoshanense gave highest amounts (6.63﹪) of chloroform extract, followed by Dendrobium loddigesii and Dendrobium falconeri, whereas, Dendrobium tosaenes and Dendrobium hoshanense yielded high quantity (27.72﹪) and (25.27﹪) by extracting with water. 2.Dendrobium crude extracts (30 μg) were screened for their ability in nitric oxide production by murine macrophage-like cell line activated with LPS and INF-γ. Dendrobium monoliforme showed high inhibitive activity of nitric oxide production whereas, Dendrobium candidum showed high productive activity of nitric oxide. 3. Cultures were grown better on B5 basal medium supplemented with 6-benzylaminopurine (BA). 4.Axillary shoots were cultured on B5 basal medium supplemented with 2.5 mg/L BA and 0.5 mg/L α-naphthaleneacetic acid (NAA) and 30 g/L sucrose under intensified light condition for 40 days. 5. The axillary shoots were also cultured on G-3 medium to obtain plantlets (4.5 cm length) for a period of 90 days. 6. Optimum medium of protocron-like bodies (PLBs) were recorded in B5 medium supplemented with 2.5 mg/L BA and 30 g/L sucrose and 3 g/L gelrite for 40 days. 7. The percentage survivals of plants were found to be 85﹪. 8. Plants were transferred to plastic cups containing a mixture of scrap of cyatheaceae, small stones (1：1) successfully acclimatized under greenhouse conditions. 9. Pharmacological studies have been performed on Dendrobium plants. 10. Dendrobium monoliforme showed better growth (based on fresh weight and dry weight basis) among the three species studies.
顯示於類別:	[藥物化學研究所] 博碩士論文

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