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| 題 名 | 利用cDNA微陣列來建構一個分子生物層級的中草藥分析平臺=Constructing a Chinese Medicine Molecular Analysis Plateform by Utilizing cDNA Microarray |
|---|---|
| 作 者 | 許志楧; | 書刊名 | 中醫藥年報 |
| 卷 期 | 25:2 2007.10[民96.10] |
| 頁 次 | 頁15-66 |
| 專 輯 | 中醫藥基因體及免疫學研究 |
| 分類號 | 414.33 |
| 關鍵詞 | 中草藥; cDNA微陣列技術; 基因表現圖譜; 標記基因; 冬蟲夏草; Chinese herbal medicine; cDNA microarray technology; Gene expression profile; Marker genes; Cordyceps sinensis; CS; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 由於醫學科技的進步,中醫藥科學化已經是時代的趨勢。然而中草藥的成份複雜,對於單味藥或傳統方劑的生產品質需要做有效的監控。另外,對於其藥理機制需要在分子生物的層次做定量的了解,以達到實驗科學的客觀驗證。為了用嚴謹的科學發法來研究中草藥,本計畫利用高效液相層析法(High Performance Liquid Chromatography, HPLC)和感應偶合電漿質譜儀(Inductively Coupled Plasma Mass Spectrometer, ICPMS),對中草藥產品品質及批次間之均一性做嚴格的監控。本計畫之目的是要利用cDNA微陣列(cDNA microarray)來建立一個分子生物層級的中草藥分析技術平臺。希望藉此計畫的完成,確立一套以科學化的方法來分析中草藥對細胞基因表現圖譜(gene expression pattern)之影響,並進而歸納出相對於該中草藥之標記基因(marker genes)。希望能據此以定量的方式,可靠且有效的了解中草藥之藥理機制。經由此技術平臺的建立,將來對各種中草藥的分析研究,皆可依循本計畫所建立之規範(包含中草藥有機/無機成份之標準化、中草藥對細胞株在分子生物層級的分析,及所應用的統計分析方法)一一加以研究。 本計畫第二年,我們完成了以下幾項目標:(1)以冬蟲夏草處理KG-1細胞後,分別在兩天、四天及八天時抽取細胞的RNA。進行螢光標示顏色對調(Dye Swap)的cDNA微陣列實驗。(2)利用統計的分析方法進行初步的基因表現圖譜分析並加以探討。(3)依據檢討結果,修正改進有機/無機化合物標準化流程,並選取兩種不同來源的天然冬蟲夏草(青海冬蟲夏草與西藏冬蟲夏草)以進行後續之分析研究。(4)以統計方法針對cDNA微陣列實驗進行迴圈設計(Loop Design)及進行基因晶片實驗。(5)以統計方法分析上述實驗結果,並初步歸納出相對於實驗性分析標的物-冬蟲夏草的標記基因。 |
| 英文摘要 | Modernization of traditional Chinese medicine has provoked great interests from bio-medical researchers globally. However, significant difficulties are associated with quality control of the multi-component herbal mixtures. Stringent quality control is the first and essential step to experimentally evaluate the pharmacological activities of traditional Chinese herbal medicine at the molecular level. HPLC (High Performance Liquid Chromatography) and ICPMS (Inductively Coupled Plasma Mass Spectrometer) were implemented as stringent quality control systems to monitor the organic- and inorganic-chemical compositions of Chinese herbal medicine. This procedure assures the uniformity between different batches of herbal mixtures for the cDNA microarray experiments. The ultimate goal of this proposal is to construct a molecular-level, analytical platform by utilizing cDNA microarray technology. As the completion of this platform, we expect to establish a standard scientific protocol for analyzing the effect of Chinese herbal medicine on cellular gene expression pattern. Thus, the potential pharmacological mechanisms could be assessed more accurately and effectively. This analytical platform, including organic- and inorganic-chemical analysis, cDNA microarray technology and statistical methods, could be applied to the investigation of a variety of different Chinese herbal medicines. In the second year of study, we have accomplished the following tasks: (1) Total RNA was extracted from KG-1 cells treated with Cordyceps sinensis for 2, 4, and 8 days. Dye swap cDNA microarray experiments were performed on these samples. (2) The patterns of global gene expression were analyzed by statistical methods. (3) Organic- and inorganic- standardization protocols were further amended according to the microarray results. Cordyceps sinensis samples of two different geological origins were chosen for the following study. (4) Loop design protocol was used for major microarray experiments. (5) Microarray results were examined by statistical methods and marker genes that are specifically regulated by Cordyceps sininsis were identified. |
本系統中英文摘要資訊取自各篇刊載內容。