川木通及常用習用品的生藥學研究
發(fā)布時間:2018-04-17 01:11
本文選題:川木通 + 生藥學鑒定 ; 參考:《西南交通大學》2017年碩士論文
【摘要】:目的:川木通系毛茛科植物小木通Clematis armandii Franch.或繡球藤Clematis montana Buch.-Ham.的干燥藤莖。具有利尿通淋,清心除煩,通經下乳的作用。目前市場混用嚴重,飲片鑒別困難,用藥安全性及藥效難以保障,本研究擬從生藥性狀、顯微、薄層定性、DNA條形碼和HPLC特征圖譜基因化學雙指紋研究,形成綜合研究體系。對市場占有率較高的川木通混品粗齒鐵線蓮和小蓑衣藤進行初步的化學成分及藥效研究,為川木通鑒定及替代品使用提供依據。方法:本次研究采用石蠟包埋技術和番紅固綠染色法對小木通、繡球藤、粗齒鐵線蓮和小蓑衣藤的莖進行了顯微鑒定,對四種藥材進行了粉末對比研究;以對照藥材為對照建立了川木通TLC鑒別方法。進行了川木通藥材的水分、總灰分以及浸出物等檢查項測定;提取生藥和飲片總DNA,對DNA的ITS2序列進行擴增測序,對DNA序列進行比較分析;采用HPLC法建立了川木通的HPLC特征圖譜,利用"中藥色譜指紋圖譜相似度評價系統(tǒng)"對收集到的樣品與小木通和繡球藤進行了相似度比對;最后為藥典法定品種與市場習用品種之間藥效區(qū)別,初步進行了藥典品種小木通、繡球藤和非藥典品種粗齒鐵線蓮和小蓑衣藤的利尿效果比較研究結果:從顯微結果看,初生射線:小木通17條,其它均為12條;繡球藤的韌皮部的厚壁細胞環(huán)帶不連續(xù)成環(huán),在髓射線處間斷;粗齒鐵線蓮呈現不明顯六凹槽,導管孔粗大;小蓑衣藤橫截面花瓣狀;粉末結果顯示,小木通、繡球藤、粗齒鐵線蓮以及小蓑衣藤在韌皮射線細胞、石細胞和導管的形態(tài)及大小上有著不同程度的差別;贗TS2的DNA條形碼結果顯示,川木通飲片主要以粗齒鐵線蓮為主,小木通使用較少,未發(fā)現繡球藤;通過NJ樹看出繡球藤、粗齒鐵線蓮和小蓑衣藤三者親緣關系相近。通過HPLC特征圖譜比對,粗齒鐵線蓮、小蓑衣藤和繡球藤在化學成分上相似度非常高;通過利尿效果研究發(fā)現粗齒鐵線蓮與川木通一樣均有明顯的利尿效果。結論:1、建立了一個川木通較為系統(tǒng)鑒定的方法,能準確有效地將川木通與鐵線蓮屬植物進行區(qū)分。2、驗證了粗齒鐵線蓮及小蓑衣藤替代川木通使用的合理性,故推薦將粗齒鐵線蓮納入藥典。
[Abstract]:Objective: Clematis armandii Franch.Or Clematis montana Buch.-Ham.A dry vine stem.With diuretic, clear heart to remove trouble, the role of menstruation under the milk.At present, the market is mixed seriously, it is difficult to identify the tablets, and the safety and efficacy of drug use are difficult to guarantee. This study aims to form a comprehensive research system from the aspects of pharmacological characters, microscopes, TLC qualitative DNA bar codes and genetic double fingerprints of HPLC characteristic maps.The chemical constituents and pharmacological effects of the mixture of Clematis caudata and Hyacinus subtilis were studied in order to provide the basis for the identification of Akebia chinensis and the use of substitute.Methods: in this study, the microscopical identification of the stems of Akebia japonicus, Hydrangea elegans, Clematis caudata and Rhizoma Hyacinus was carried out by paraffin embedding technique and red solid green staining, and the powder comparison of the four medicinal materials was carried out.A TLC method for identification of Akebia chinensis was established.The contents of water, total ash and extracts of Akebia chinensis were determined, the ITS2 sequence of DNA was amplified and sequenced, and the DNA sequence was compared and analyzed. The HPLC characteristic map of Akebia chinensis was established by HPLC method.The similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine was used to compare the similarity between the collected samples and Acacia chinensis and Hydrangea elegans.A comparative study of diuretic effects of the pharmacopoeia varieties of Akebia, Hydrangea and non-pharmacopoeia varieties was carried out. The results were as follows: from the microscopic results, the primary rays were 17 and 12 of the others;The thick-walled cell band of phloem of Hydrangea tenuifolia is discontinuous and discontinuous in the medullary ray; the clematis has no obvious six grooves, the tube hole is thick; the cross section of the small coir coat is petal-like; the powder results show that the small Akebia, Hydrangea,The morphology and size of phloem ray cells, stone cells and ducts were different from each other.The results of DNA bar code based on ITS2 showed that the main components of Akebia chinensis were clematis, but the use of Akebia was less, and no Hydrangea was found, and the genetic relationship among them was similar by NJ tree.The comparison of HPLC characteristic maps showed that the similarity of chemical constituents of Clematis caudata and Hyacinth was very high, and the diuretic effect of Clematis crassifolia and Akebia chinensis was found to be the same as that of Akebia chinensis.Conclusion: 1. A systematic identification method for Akebia chinensis was established, which could distinguish Akebia from Clematis accurately and effectively.Therefore, it is recommended that the coarse-toothed clematis into the pharmacopoeia.
【學位授予單位】:西南交通大學
【學位級別】:碩士
【學位授予年份】:2017
【分類號】:S567
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