魚針草(Anisomelesindica(L.)Kuntze)基原鑑定、指標成分純化分析及系列化學成分生理活性綜論

Studies on the original plants identification, biological activities and analysis of marker compounds from Anisomeles indica (L.) KUNTZE

李雅婷(Ya-Ting Lee)

指導教授 ： 曾耀銘

朝陽科技大學/理工學院/生物技術與化學工程研究所碩士班/碩士(2010年)

摘要

魚針草 ( Anisomeles indica (L.) KUNTZE ) 始載於生草藥性備要，為唇形科(Labiatae)之一年或越年生草本植物，近代研究顯示全草具有解熱、袪風、除濕、健胃、解毒、止痛、抗菌之功效。一般民間常用於治療風濕性關節炎、腸胃炎及其它炎症等。本實驗研究分為兩階段：首先以傳統生藥組織切片，比較藥材形態相近於魚針草之豨薟草(Siegesbeckia orientalis (L))與魚針草內部構造之差異，並利用核糖體內轉錄區間(Internal Transcribed Spacer；ITS)序列定序，鑑定魚針草之基原，並比較魚針草與豨薟DNA序列上之差異；ITS序列定序能提供客觀的依據，輔佐形態學鑑識主觀上之不足；而後取魚針草乾燥全草以甲醇熱迴流萃取，再將甲醇萃取濃縮物以極性分配為氯仿層、正丁醇層與水層，在正丁醇層以矽膠管柱層析、薄膜層析、再結晶、製備型高效能液相層析儀進行化合物分離。
研究結果顯示，針對魚針草與豨薟藥材的外觀形態特徵與內部組織構造進行剖析，建立標準生藥組織圖，，以便於比對魚針草與豨薟藥用植物於市場上真偽之情況；在魚針草與豨薟ITS序列相似程度低差異大，明顯比較出其基原之不同；目前於NCBI上魚針草之ITS序列並無人發表，而本研究提供魚針草之ITS序列，可供他人做研究比對之參考，而在化學指標成分分析鑑定之實驗結果得到ovatodiolide與其含量較多之3個化合物，該3個化合物經各種光譜分析結構鑑定之證實為acteoside、β-sitosterol、stigmasterol。
利用組織顯微鑑定法、指標成分分析及分子生物鑑定法進行其鑑別之研究，釐清其基原，建立科學化的鑑別機制，以供未來鑑別之依據，能更快速及準確的比對正確之基原，並建立魚針草指標性成分的定量分析方法，期望魚針草未來成為現代科學化中藥及保健食品與新藥開發之潛力。

關鍵字

內轉錄區間 ； 基原鑑定 ； 魚針草內酯 ； 洋丁香? ； 魚針草

並列摘要

Anisomeles indica (L.) KUNTZE. (Labiatae) is an annual and biennial herb. In an alternative medicine this plant has been used as damp heat, carminative, stomachic, detoxification, pain relief, rheumatoid arthritis, gastroenteritis and other inflammatory diseases. It is one of the popular Traditional Chinese Medicine (TCM).
In the present study, we firstly started with literature reviews on Pentsaology, Pharmacognosy and Pharmacology of A. indica. Then, we collected specimen of these two indigenous species for tissue identification. we also analyzed the ITS (Internal Transcribed Spacers) sequences of the two species. According to the result, these medicinal plant samples were identified as A. indica and S. orientalis .
The stem of A. indica were extracted with methanol under reflux. The combined methanol extract was portioned with chloroform and n-butanol. Each layer was concentrated under reduced pressure individually, and then subjected to silica gel column chromatography, thin-layer chromatography, and preparative HPLC. Four compounds were isolated among 4 compounds structures were also determined. The four compounds namely：acteoside、ovatodiolide、β-sitosterol、stigmasterol.
We analyzed the content of indicator ingredients of acteoside and ovatodiolide in these samples with HPLC method. We also analyzed the ITS (Internal Transcribed Spacers) sequences of the two species.
Through this study, we expect to initiate an identification system for the local origins of A. indica in Taiwan and explore its potential usages as a native medicine resource in the future.

並列關鍵字

acteoside ； ovatodiolide ； identification ； internal transcribed spacer ； Anisomeles indica

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