陈涛,何磊,林燕
CHEN Tao,HE Lei,LIN Yan

• 1:贵州医科大学药学院
• 2:贵州医科大学

摘要(Abstract)：

目的 探讨苗药头花蓼生药综合鉴定的方法。方法 选取黔产苗药头花蓼药材10批次，制作头花蓼药材植物标本，对其根、茎、叶、花、果等形态特征进行性状鉴别;制作头花蓼药材粉末，分别采用氯化铁(FeCl_3)显色反应、盐酸-镁粉反应、氯化铝(AlCl_3)显色反应及薄层色谱法进行理化鉴别;采用显微镜对其药材粉末进行显微鉴别;采用植物基因组DNA提取试剂盒进行DNA分子标记鉴定。结果 头花蓼茎丛生，叶椭圆形、长2～5 cm、宽1～2 cm、叶先端急尖基部楔形，花序头状、单生或顶生、成对，总状花序直立、近球形，花被淡红色;苗药头花蓼醇提液在FeCl_3显色反应中显蓝色、久置变深蓝色，在盐酸-镁粉反应中显深红色，在AlCl_3显色反应中于360 nm紫外灯下显黄色荧光;头花蓼粉末显微鉴别具有花粉粒、晶鞘纤维、纤维素、导管、淀粉粒、木栓细胞等特征碎片;头花蓼药材在500 bp附近显示清晰明亮的单一条带，测序显示序列差异主要集中在ITS1和ITS2区。结论 由性状鉴别、理化鉴别、粉末显微鉴别及DNA分子标记鉴定结果，阐明了原鉴定药材为蓼科蓼属植物头花蓼Polygonum capitatum Buch.-Ham. ex D. Don。
Objective To investigate the systematic evaluation of pharmacognostic identification of Polygonum capitatum. Methods Ten batches of P. capitatum cultivated in Guizhou were chosen for plant samples. Macroscopical identification was conducted on plant roots,stem,leave,flower and fruit. The P. capitatum powder was processed for physical and chemical distinction by FeCl_3 chromogenic reaction, hydrochloric acid magnesium powder reaction, AlCl_3 color development reaction,and thin-layer chromatography. Microscope identification was carried out on the powder.Plant genome DNeasy Plant Kit was adopted to identify DNA molecular marker identification.Results Theresults showed that the stem of P. capitatum was tufted,the leaves were oval,2 to 5 cm long,and 1 to 2 cm wide; the leaf apex was acute and cuneate at the base,the inflorescence was capitate,paired or solitary; the raceme was erect and nearly spherical,and the perianth was light red.Furthermore,for the chromogenic reaction of FeCl_3 ethanol extract of P. capitatum,appeared blue and turned to dark blue after long time storing at room temperature. For the reaction of hydrochloric acid magnesium powder,the alcohol extract of P. capitatum,exhibited deep red. In the color reaction of AlCl_3,the alcohol extract revealed yellow fluorescence under 360 nm UV lamp. Microscope identification of the powder displayed pollen grains,crystal sheath fibers,cellulose,vessels,starch grains,cork cells,and other characteristic fragments. In addition,DNA barcoding electrophoresis results showed that P. capitatum showed a clear and bright single band near 500 bp,and further sequencing results showed that the sequence differences were mainly concentrated in ITS1 and ITS2region. Conclusion Systematic evaluation for the identification of P. capitatum is established,which combines with macroscopical identification, physicochemical identification, powder microscope identification,and DNA molecular identification. Finally,the original medicinal material is identified as P. capitatum Buch-Ham. ex D. Don.

关键词(KeyWords)： 中草药;头花蓼;生药鉴定;性状鉴别;理化鉴别;显微鉴别;DNA分子标记鉴定
Chinese herbal drugs;Polygonum capitatum (P. capitatum);pharmacognostic identification;macroscopical identification;physicochemical identification;microscope identification;DNA molecular identification

Abstract:

Keywords:

基金项目(Foundation): 贵州省卫生健康委科学技术基金(gzwkj2021-449);; 贵州省科技计划项目(黔科合基础-ZK[2022]一般362);; 全国大学生创新训练计划项目(202210660131)

作者(Author): 陈涛,何磊,林燕
CHEN Tao,HE Lei,LIN Yan

参考文献(References)：

• [1]贵州省药品监督管理局．《贵州省中药材、民族药材质量标准》[M]．贵阳:贵州科技出版社，2003:68-69．
• [2]国家中医药管理局《中华本草》编委会．中华本草·苗药卷[M]．贵阳:贵州科技出版社，2005:223-224．
• [3]HE C Y,FU J,MA J Y,et al.Biotransformation and in vitro metabolic profile of bioactive extracts from a traditional miao-nationality herbal medicine,polygonum capitatum[J].Molecules,2014,19(7):10291-10308．
• [4]LIAO S G,ZHANG L J,SUN F,et al.Antibacterial and anti-inflammatory effects of extracts and fractions from polygonum capitatum[J].J Ethnopharmacol,2011,134:1006-1009．
• [5]HUANG Y,ZHOU Z Y,YANG W,et al.Comparative pharmacokinetics of gallic acid,protocatechuic acid,and quercitrin in normal and pyelonephritis rats after oral administration of a Polygonum capitatum extract[J].Molecules,2019,24:3873．
• [6]张丽艳，刘昌孝，唐靖雯，等．头花蓼提取物中具抗淋球菌作用的有效部位研究[J]．中草药，2019,50(2):177-181．
• [7]尹艳，赵德刚，党伯岳，等．头花蓼优质种源筛选及其四倍体质量评价研究[J]．种子，2015,34(11):63-65．
• [8]吕炎唏，王隶书，程东岩，等．中药头花蓼的化学成分和药理作用研究概况[J]．中国药师，2017,20(10):1849-1853．
• [9]LIN Y,HE L,CHENG X J,et al.Polygonum capitatum,the hmong medicinal flora:a comprehensive review of its phytochemical,pharmacological and pharmacokinetic characteristics[J].Molecules,2022,27,6407．
• [10]郑林，陈浩，孙慧园，等．头花蓼提取物在肾盂肾炎模型与正常大鼠体内的组织分布比较研究[J]．中国新药杂志，2019,28(8):1008-1016．
• [11]张梦薇，罗昭逊，孙朝琴，等．头花蓼灌胃对大鼠幽门螺杆菌相关性胃炎的治疗作用及其机制[J]．山东医药，2018,58(15):35-38．
• [12]孙长生，梁斌，王传芳．头花蓼研究进展[J]．中国现代中药，2005,7(4):26-28．
• [13]周涛，谢宇，张丽艳，等．基于SCAR分子标记的头花蓼分子鉴定研究[J]．中国中药杂志，2013,38(16),2577-2580．
• [14]OLIVEIRA R R M,NUNES G L,DELTG L,et al.PIPE-BAR and Overlap PER:tools for a fast and accurate DNAbarcoding analysis and paired-end assembly[J].BMCbioinformatics,2018,19(1):297．
• [15]杨倩倩，刘苏汶，俞晓平．DNA条形码分析方法研究进展[J]．应用生态学报，2018,29(3):1006-1014．
• [16]FANG R Z,LI X F,KONG J,et al.Anti-inflammatory effect and pharmacological study of polygonum capitatum based on network pharmacology[J].Earth Environmental Science,2021,632:032013．
• [17]YANG Y,YANG Y B,LU W Q,et al.A new hydroxyjasmonic acid derivative from polygonum capitatum[J].Chem Nat Compd,2017,53(3):417-421．
• [18]刘慧娟．三氯化铁试剂在炔醇衍生物合成中的应用[D]．厦门:厦门大学，2007．
• [19]桂利利，吴正坤，余惠凡，等．黄药叶总黄酮提取工艺优化及其抗氧化活性研究[J]．食品研究与开发，2021,42(19):100-108．
• [20]龚元，李咏梅，陈明．基于盐酸-镁粉反应比色法的梭洞学中总黄酮含量的测定[J]．安徽农业科学，2010,38(11):5636-5637．
• [21]杨志欣，王海威，张文君，等．苦参总黄酮含量测定方法的优化[J]．中成药，2017,39(5):946-951．
• [22]张国权，罗崧方，黄建蓉，等．三氯化铝比色法检测桑叶提取物总黄酮的研究[J]．农产品加工，2018(7):52-57．
• [23]刘云鹤，王晓林，钟方丽．三氯化铝显色法测定刺玫叶提取物总黄酮的含量[J]．特产研究，2017,39(2):50-53．
• [24]石乃星，文国松，赵明富．黄精属植物DNA分子鉴定技术应用研究进展[J]．植物遗传资源学报，2021,22(5):1209-1218．
• [25]王刚，曹佩，韦学敏，等．分子标记技术在药用植物种质资源研究中的应用[J]．中国现代中药，2019,21(11):1435-1444.

文章评论(Comment)：