2013-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/654398摘要：根據民國 99年行政院衛生署十大癌症死因統計，肝癌於不分男女之排名中佔癌症主要死因第二位，更高於全世界所排名之第四位，因此在台灣如何維護肝臟健康是值得重視的議題。近年研究發現，牛樟芝具有護肝、抗發炎、抑癌等生理活性，然而對牛樟芝子實體「活性成分」之研究則仍有很大的空間待加強。目前牛樟芝人工栽培方式有液態發酵、固態培養、椴木栽培三種，而其中三&#33820;類與固醇類含量以椴木栽培較高，故本實驗室特別以「椴木栽培」之牛樟芝子實體作為實驗材料，從中萃取出多種重要之純化物，且已在三&#33820;類純化物對抗肝癌研究獲得良好的成果，並藉此建立出細胞死亡模式之「系統分析平台」。然而，牛樟芝子實體中麥角固醇含量甚多，且具有很高的抑癌潛力，因此本計畫將以「麥角固醇」純化物做進一步的研究，擬分兩年完成，計畫細部內容如下： 第一年：從牛樟芝萃取出含量較高的四種重要麥角固醇純化物：ergostatrien-3β-ol、zhankuic acid A、antcin C、zhankuic acid C，評估此等抑制肝癌細胞增殖之效果，並探討其對正常肝細胞生長之影響，以篩選出具較高抗肝癌潛力之牛樟芝麥角固醇純化物，更進一步以本實驗室已建立的細胞死亡模式之「系統分析平台」進行細胞死亡模式先期評估。 第二年：更深入以細胞死亡模式之「系統分析平台」分析其所誘導之細胞死亡模式，包括進行細胞週期分析與雙染分析以測定「細胞凋亡」；檢測乳酸脫氫&#37238;之釋放，並使用流式細胞儀進行前向、側向散色光測量以測定「細胞壞死」；藉由免疫螢光染色觀察細胞之自噬體生成以測定「自體吞噬」，並且更深入探討此等細胞死亡模式之相關分子機制。最後，進一步評估其抑制肝癌細胞轉移之功效，包含移行、侵犯、黏附能力試驗，以作為牛樟芝子實體發展為保健食品和輔助抗癌治療之參考。<br> Abstract: According to the statistics in 2010,liver cancer is the second leading cause of cancer related deaths in Taiwan,which is higher than the forth in the world.Therefore,how to keep the liver healthy is a big issue.Recent researches have established that Antrodia cinnamomea,a Taiwan specific medicinal mushroom can manipulate biological activities including hepatoprotection,anti-inflammation and anti-cancer activities.However,research focus on its “active compounds” is limited. Liquid-state fermentation,solid-state culture,and basswood cultivation,are the widely adopted methods for artificial cultivation of Antrodia cinnamomea and among these three cultivation methods, basswood cultivation can obtain higher quantities of the functional ingredients.Hence our laboratory employed Antrodia cinnamomea harvested from basswood cultivation to extract variety of purifications.Our recent studies have analysed and established a cell death model and the purified triterpenoids exhibited a good result of anti-hepatoma activity.In addition,ergosterols which are abundant in Antrodia cinnamomea,have potential function of anti-cancer.Consequently,the ergosterols from Antrodia cinnamomea will be used for further study: First year: Initially the predominantly present four ergosterols:ergostatrein-3β-ol,zhankuic acid,antcin C and zhankuic acid C,which are extracted from Antrodia cinnamomea will be evaluated for their efficacy in inhibiting hepatocellular carcinoma proliferation and their effect on normal hepatic cell growth.Based on this preliminary screening,the best promising ergosterol with the highest potential of anti-hepatoma will be selected for in-depth study. Second year: The mechanism of action of this screened ergosterol will be elucidated and the cell death models, including apoptosis,necrosis,autophagy will be investigated further.Finally,this project will evaluate the anti-hepatocellular carcinoma metastasis ability of ergosterols by migration,invasion and adhesion assay.Hopefully,this identified ergosterol with anti-hepatoma activity from the fruiting body will be applied in the development of Health Food and the adjuvant therapy of cancer.肝癌椴木栽培牛樟芝子實體麥角固醇自體吞噬liver cancerbasswood cultivationthe fruiting body of Antrodia cinnamomeergosterolsautophagy