利用生物冷光分析技術建立中草藥快速品質檢驗平台系統

A Quality Control Plateform for Fast Detecting Chinese Herbal Medicines by Using Cell-Based Bio-luminesence Analysis

張芷瑜(Chih-Yu Chang)

指導教授 ： 吳瑞裕

臺北醫學大學/醫學院/醫學科學研究所/碩士(2010年)

https://doi.org/10.6831/TMU.2010.00058

摘要

傳統的中草藥鑑別與品質管制是目前中藥科學化最為關鍵，而且是發展瓶頸的最主要原因。大多是由其外觀、氣味、顏色加以辨識，進一步者會利用植物組織上的特徵，以組織切片或粉末進行顯微鏡檢驗。但是當藥材因為加工處理後其外觀無法辨識時，傳統鑑別方法便無法派上用場。近年來隨著分子生物技術的進步，利用DNA序列的分析技術，觀察生物遺傳物質的特徵，但此技術雖能準確區別藥材間基原的異同，卻不一定能直接反映其中化學成分的變化；而中草藥的品質可能因為產地、採收季節與生長時間而影響到其中的眾多化學成分，目前應用較廣的鑑定方式為利用高效液相層析儀( HPLC )建立中草藥化學指紋圖譜，以分析藥材中的化學成分，將特定的化合物視為某藥材的特徵性物質，然而，現今在中草藥的使用上多為複方，且單一種中草藥中便含有多種化學成分，若以特定的化合物作為藥材的指標性成分，仍無法實際的反應出中草藥在生物體內的真正療效，且由於上述的方式過程繁複且所需時間長，無法真正的因應中草藥品種鑑定與品質管理實際需求。因此，本研究計畫為能實際觀察中草藥對於生物體所產生的作用，利用生物感測器的模式，發展出一套快速的中草藥檢驗帄台系統，透過大腸桿菌( E. coli )中啟動子針對不同中草藥的不同反應作為中草藥品種及品質的鑑定。本實驗利用基因轉殖技術，將31種啟動子( promoter )個別與帶有冷光基因( luxCDABE )之無啟動子質體( promoterless vector )進行轉殖，再與大腸桿菌進行轉形，將轉形後的菌種加入中藥萃取物或是其指標成分，藉由各類中草藥對於不同啟動子( promoter )產生直接或間接的影響，造成大腸桿菌所產生之生物冷光的變化不同，以此建立各類中草藥萃取物固定的冷光圖譜，進而快速檢驗中草藥的來源及種類。

關鍵字

中草藥 ； 啟動子 ； 生物感測器 ； 報導基因 ； 冷光基因 ； 品質鑑定 ； 快速檢驗平台

並列摘要

Traditional Chinese medicine played an important role in the health of the Chinese people for thousands of years. Herbal medicines and their derived products are widely used as therapeutic products in many countries. Most of herbal medicines and their derivative products were often prepared from the crude plant extracts, which comprise a complex mixture of different chemical constituents. The chemical compounds of these constituents are different from the different species. However, the same drug from the same plant species may contain different contents of the chemical constituents, the degree of difference may depends on various factors such as often cited geographic location, season and time of harvest, post-harvest handling, drying process and storage. Therefore, the quality control of the herbal medicines and their derived products is difficult. The traditional identifying method of herbal medicines is according to the appearance of plant. Currently, there are various modern techniques available such as genetic fingerprinting and chromatographic fingerprinting of the chemical component, especially by high-performance liquid chromatography( HPLC ). Nevertheless, these analysis methods of one or several active components in a herbal medicine does not represent its quality because its curative efficacy results from multiple components at multiple targets within an organism. In this study, to observe the reaction of herbal medicine within an organism practically, we have constructed 31 kinds of plasmids with one promoterless bacterial luciferase genes, luxCDABE, from Photorhabdus luminescens and one promoter each. Expression of each luxCDABE genes was driven by 31 different promoters separately. The effects of herbal medicines and their derived products to 31 clones will increase or lower the expression of bioluminescence of E. coli. Therefore, a bioluminescence technology for quality control of traditional Chinese medicine was established.

並列關鍵字

chinese herbal medicine ； biosensor ； reporter gene ； promoter ； luxCDABE operon ； quality control

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