雷公藤药材基原考证及分子鉴定研究

Textual research on origin and molecular identification of Tripterygii Radix

目的为确保雷公藤药材临床用药的安全性和有效性,对雷公藤药材基原进行文献考证,并建立特异性快速聚合酶链式反应(PCR)分子鉴定方法。方法查阅文献对雷公藤基原进行考证;对雷公藤及混伪品DNA条形码进行对比分析,筛选具有稳定差异的特异性鉴别位点,设计鉴别引物,优选出特异性快速PCR扩增条件。结果文献考证发现雷公藤和昆明山海棠在植物形态特征、药用部位次生代谢产物特征及遗传信息系统分化特征上存在大量中间类型,种间界限模糊。建立可用于鉴定雷公藤药材真伪的特异性快速PCR方法,所有正品雷公藤在300~400 bp间均能扩增出目的基因条带,混伪品则无对应条带。结论建议将雷公藤与昆明山海棠二者合并为雷公藤药材来源;建立了快速、准确、稳定的特异性快速PCR方法,可用于雷公藤药材与其混伪品的真实性鉴定。

Objective To ensure the safety and efficacy of Tripterygii Radix for clinical use,we conducted a literature review on the origin of Tripterygii Radix and established a specific rapid PCR molecular identification method.Methods The origin of Tripterygii Radix were validated by reviewing literature reports;the DNA barcodes of Tripterygii Radix and the forgeries were compared.The specific identification sites with stable differences were screened,the identification primers were designed and the specific rapid PCR amplification conditions were optimized.Results The literature review revealed that Tripterygium wilfordii and Tripterygium hypoglaucum hutch had a large number of intermediate types and blurred interspecific boundaries in terms of plant morphological characteristics,secondary metabolite characteristics of medicinal parts and genetic information system differentiation.A specific rapid PCR method was developed to identify the authenticity of Tripterygii radix.All authentic Tripterygii Radix were able to amplify the target gene bands between 300 and 400 bp,while no corresponding bands were found for the forgeries.Conclusion It is suggested that Tripterygium wilfordii and Tripterygium hypoglaucum hutch should be combined as a source of Tripterygii Radix,and a rapid,accurate and stable specific rapid PCR method should be established for Tripterygii Radix and its hybrid for authenticity identification.