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題 名 | 樟芝栽培與其抗癌活性之研究=Study on the Culture Techniques and Anti-Tumor Activities of Antrodia Cinnamomea |
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作 者 | 張懿欣; | 書刊名 | 中醫藥年報 |
卷 期 | 29:5 2011.09[民100.09] |
頁 次 | 頁1-32 |
專 輯 | 傳統藥材生技之研究 |
分類號 | 414.34 |
關鍵詞 | 牛樟芝; 人工栽培; 樟芝萃取液; 抗癌活性成分分析; Antrodia cinnamomea; Anti-tumor activities; DNA fragmentation; |
語 文 | 中文(Chinese) |
中文摘要 | 牛樟芝(Antrodia cinnamomea)是臺灣特有褐腐菌,只能生長在臺灣常綠闊葉樹-牛樟(Cinnamomum kanehirae)的內壁。牛樟芝氣味芳香,味辛苦,子實體富含多醣體與三萜類等成分,有抗B 型肝炎與防癌等功效。基於其功效以及野生牛樟芝子實體數量極少,加上人工栽培不易等因素,所以價格昂貴;業者往往為了採取野生牛樟芝而大肆盜伐牛樟樹,使臺灣特有珍貴樹種瀕臨滅絕,破壞森林多樣性與生態平衡。本實驗室之研究以建立藥用牛樟芝人工培養技術及篩選並應用研發藥用牛樟芝成分抗癌活性相關產品為主要研究目的。第一階段以嘉義大學林產科學系與森林暨自然資源學系為主,研發並建立最佳之牛樟芝人工培養條件與技術。第二年則由中山醫學大學和弘光科技大學負責,以醫學生物角度出發,分析鑑定第一階段實驗室之人工栽培牛樟芝抗癌活性功效的成分。此外,本實驗室也探討靈芝與與黑木耳萃取液之抗癌活性,並比較此兩種菇類與牛樟芝之抗癌活性。研究結果顯示,最佳樟芝菌絲體人工培養基為含有8 g/L 瓊脂濃度之MPA 配方;而最佳之培養生長條件為中性或弱酸性pH 值,培養溫度為30℃。本實驗室並首度發現添加米糠可有效促進樟芝在太空包培育中的生長,而且樟芝可在含有相思樹配方的太空包基質中形成原質。本實驗室利用細胞生長曲線、DNA 斷裂分析與癌細胞在半固態培養基上的群落分析等研究法多方面探討人工栽培樟芝的功效。樟芝發酵液、樟芝水萃物與樟芝甲醇萃取物都可抑制人類H1299 肺癌細胞株、HepG2 肝癌細胞株與CaCO2 大腸結腸癌細胞株的生長;但以樟芝發酵液抑制癌細胞生長的效果較佳。抑制癌細胞生長之活性比較,以樟芝發酵液的抗癌功效最佳,其次為樟芝甲醇萃取物、靈芝水萃物、黑木耳水萃物與樟芝水萃物。本實驗室也利用DNA 斷裂分析探討上述萃取物是否可誘發癌細胞之細胞自戕作用(apoptosis),達到抑制癌細胞生長的效果。實驗結果顯示樟芝發酵液、樟芝水萃物與樟芝甲醇萃取物都可以使H1299、HepG2 與CaCO2 之DNA 產生斷裂,其中又以樟芝水萃物與甲醇萃取物誘發癌細胞DNA 斷裂的情形較為顯著。而癌細胞在半固態培養基的群落分析方面,H1299 在含有樟芝發酵液、樟芝水萃物與樟芝甲醇萃取的半固態培養基中,隨著發酵液與萃取物濃度增高,細胞株群落形成數隨之遞減;但高濃度樟芝發酵液與樟芝水萃物可完全抑制H1299 在半固態培養基中形成細胞株群落。綜合本實驗室上述實驗結果,樟芝發酵液抑制癌細胞生長功效較佳;由DNA 斷裂分析得知樟芝水萃物與甲醇萃取物則可較有效地誘發癌細胞之細胞自戕作用,而樟芝發酵液與樟芝水萃物則可有效抑制癌細胞在半固態培養基中形成細胞株群落的活性。本實驗室之研究成果不但已建立牛樟芝的最佳人工栽培條件與技術,並已證實在實驗室利用人工栽培之牛樟芝具有抗腫瘤細胞生長與誘發細胞自戕作用的活性。未來本實驗室將以上述實驗結果為基礎,進一步探討人工栽培牛樟芝抗癌活性與增進人體免疫作用之作用機制,同時也將分析其活性成分,研發相關之抗癌保健產品。 |
英文摘要 | Antrodia cinnamomea is a unique brown-rot fungus species which only parasitizes in the decayed heart wood of stout camphor (Cinnamomum kanehirae) in Taiwan. The fruiting body of A. cinnamomea contains abundant polysaccharrides and terpenes that are proved to have effective anti-viral function and anti-tumor activity. The price of A. cinnamomea therefore rocketed in the market and it has become the target of illegal harvesting. Consequently, C. kanehirae has become an endangered species in Taiwan due to the illegal felling. The present proposal is divided into 2 stages: the first stage is aimed to develop optimal culture conditions and techniques for cultivating A. cinnamomea. The second stage is to identify and compare the effective and active anti-tumor component(s) of cultured A. cinnamomea, and commercial available Ganoderma and Auricularia by the first stage. The active components and the ratio of active metabolites of A. cinnamomea derived from Lab. culture will be further analyzed. The purpose of this study was to investigate the anti-tumor components from A. cinnamomea to induce apoptosis in human liver, lung and intestine cancer cells. In addition to compare the anti-tumor components of cultured A. cinnamomea with Ganoderma and Auricularia. The results showed that the MPA we used was containing 8 g/L agar, and cultured in the 30℃, neutral or weak-acid condition. Mycelium could be promoted growth by rice bran and be formed the mycelium substrate in the space package which containing the prescription of Acacia confuse. Treatment of H1299, HepG2 and CaCO2 cells with A. cinnamomea extracts resulted in dose and time-dependent sequences of events marked by apoptosis, as evidenced by cell viability, DNA fragmentation and the assay of clonogenicity. The results showed that the fermented liquid, water and methanol-extracted A. cinnamomea can suppress the growth of the cancer cells. Our finding suggested that fermented liquid of A. cinnamomea have better anti-tumor effect than methanol-extracted A. cinnamomea,water-extracted Ganoderma, water-extracted Auricularia and water-extracted A. cinnamomea. In DNA fragmentation assay, water-extracted and methanol-extracted A. cinnamomea have significant expression. Treatment of H1299 cell with A. cinnamomea extracts resulted in colony inhibition which was accompanied by dose-dependent manner. Analysis of the study data suggests that We not only develop optimal culture conditions and techniques for cultivating A. cinnamomea, but also demonstrate the A. cinnamomea extracts suppress the growth of the cancer cells and induce cell apoptosis. It may possess anti-tumor properties potentially valuable for application in health products. |
本系統中英文摘要資訊取自各篇刊載內容。