川木通及常用习用品的生药学研究

发布时间：2018-04-17 01:11

本文选题：川木通 + 生药学鉴定　；参考：《西南交通大学》2017年硕士论文

【摘要】：目的:川木通系毛茛科植物小木通Clematis armandii Franch.或绣球藤Clematis montana Buch.-Ham.的干燥藤茎。具有利尿通淋,清心除烦,通经下乳的作用。目前市场混用严重,饮片鉴别困难,用药安全性及药效难以保障,本研究拟从生药性状、显微、薄层定性、DNA条形码和HPLC特征图谱基因化学双指纹研究,形成综合研究体系。对市场占有率较高的川木通混品粗齿铁线莲和小蓑衣藤进行初步的化学成分及药效研究,为川木通鉴定及替代品使用提供依据。方法:本次研究采用石蜡包埋技术和番红固绿染色法对小木通、绣球藤、粗齿铁线莲和小蓑衣藤的茎进行了显微鉴定,对四种药材进行了粉末对比研究;以对照药材为对照建立了川木通TLC鉴别方法。进行了川木通药材的水分、总灰分以及浸出物等检查项测定;提取生药和饮片总DNA,对DNA的ITS2序列进行扩增测序,对DNA序列进行比较分析;采用HPLC法建立了川木通的HPLC特征图谱,利用"中药色谱指纹图谱相似度评价系统"对收集到的样品与小木通和绣球藤进行了相似度比对;最后为药典法定品种与市场习用品种之间药效区别,初步进行了药典品种小木通、绣球藤和非药典品种粗齿铁线莲和小蓑衣藤的利尿效果比较研究结果:从显微结果看,初生射线:小木通17条,其它均为12条;绣球藤的韧皮部的厚壁细胞环带不连续成环,在髓射线处间断;粗齿铁线莲呈现不明显六凹槽,导管孔粗大;小蓑衣藤横截面花瓣状;粉末结果显示,小木通、绣球藤、粗齿铁线莲以及小蓑衣藤在韧皮射线细胞、石细胞和导管的形态及大小上有着不同程度的差别。基于ITS2的DNA条形码结果显示,川木通饮片主要以粗齿铁线莲为主,小木通使用较少,未发现绣球藤;通过NJ树看出绣球藤、粗齿铁线莲和小蓑衣藤三者亲缘关系相近。通过HPLC特征图谱比对,粗齿铁线莲、小蓑衣藤和绣球藤在化学成分上相似度非常高;通过利尿效果研究发现粗齿铁线莲与川木通一样均有明显的利尿效果。结论:1、建立了一个川木通较为系统鉴定的方法,能准确有效地将川木通与铁线莲属植物进行区分。2、验证了粗齿铁线莲及小蓑衣藤替代川木通使用的合理性,故推荐将粗齿铁线莲纳入药典。
[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.
【学位授予单位】：西南交通大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567

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