四川盆周山地重楼属植物药用资源的筛选及评价
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摘要
目的挖掘具有药用价值的优异重楼属植物资源,缓解药用重楼资源压力,为促进重楼的可持续利用提供材料基础。方法在四川盆周山地采集21份重楼资源,根据表型筛选出大块茎资源GQ与MH,多芽资源GK。初步形态鉴定后,利用DNA条形码ITS、psbA-trnH序列,采用Kimura双参数模型构建ML系统聚类树,进行系统发育分析。采用HPLC法检测其主要活性成分皂苷的含量,以预测其潜在药用价值。结果形态学结合分子鉴定结果表明,GQ、GK和MH分别依次为华重楼、滇重楼和长柱重楼。GQ、GK、MH和对照滇重楼(CK)的重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的含量及其比例表现各异,其中总含量表现为GQ>MH>CK>GK,重楼皂苷Ⅰ在GQ和MH中占比较高,分别较CK高出67.61%和73.25%,而重楼皂苷Ⅶ在GK中占比最高,表现为56.38%。结论 GQ、GK和MH分别为产于四川盆周山地特定居群的华重楼、滇重楼和长柱重楼,引种至成都平原后性状表现优良,为后续的重楼的育种研究提供了材料基础;同时3个资源均具药用潜力,其中长柱重楼MH居群资源的重楼皂苷Ⅰ及重楼皂苷总含量较高,为滇重楼、华重楼替代药材的筛选提供了新选择。
Objective Mining Paris resources with medicinal value could underlay the breeding selection and relieve the pressure on wild resources.Methods Twenty-one Paris resources in the mountain area of Sichuan Basin were collected and selected according to their phenotype and rhizome features.Two resources GQ,MH with bigger rhizome and one resource(GK) with polygemmic feature were screened.After preliminary identification,based on Kimura-2-parameter model,molecular phylogenetic trees were constructed based on the different sequence of ITS and psbA-trnH between the screened resources and the homologous sequences from NCBI using the MaximumLike(ML) method.Main active saponin was determinated by HPLC method to predict its potential medicinal value.Results GQ,GK,and MH were special resources of P.polyphylla var.chinensis,P.polyphylla var.yunnanensis,and P.forrestii,respectively,in mountains around Sichuan Basin.The content and proportion of polyphyllin Ⅰ,Ⅱ,Ⅵ,Ⅶ in GQ,GK,MH were different.The total content was GQ>MH>CK>GK.The proportion of polyphyllin Ⅰ in GQ and MH was 67.61% and 73.25% higher than CK,respectively.While the proportion of polyphyllin Ⅶ was most in GK(56.38%).Conclusion This study specified three Sichuan local Paris resources with excellent rhizome features.And they performed well after introduced to Chengdu Plain providing the material basis for the follow-up breeding study of Paris.Three resources have medicinal potential,especially the polyphyllin Ⅰ and polyphyllin content in MH(P.forrestii) is higher,which can provide a new choice for screening substitution materials of P.polyphylla var.chinensis and P.polyphylla var.yunnanensis.
Objective Mining Paris resources with medicinal value could underlay the breeding selection and relieve the pressure on wild resources.Methods Twenty-one Paris resources in the mountain area of Sichuan Basin were collected and selected according to their phenotype and rhizome features.Two resources GQ,MH with bigger rhizome and one resource(GK) with polygemmic feature were screened.After preliminary identification,based on Kimura-2-parameter model,molecular phylogenetic trees were constructed based on the different sequence of ITS and psbA-trnH between the screened resources and the homologous sequences from NCBI using the MaximumLike(ML) method.Main active saponin was determinated by HPLC method to predict its potential medicinal value.Results GQ,GK,and MH were special resources of P.polyphylla var.chinensis,P.polyphylla var.yunnanensis,and P.forrestii,respectively,in mountains around Sichuan Basin.The content and proportion of polyphyllin Ⅰ,Ⅱ,Ⅵ,Ⅶ in GQ,GK,MH were different.The total content was GQ>MH>CK>GK.The proportion of polyphyllin Ⅰ in GQ and MH was 67.61% and 73.25% higher than CK,respectively.While the proportion of polyphyllin Ⅶ was most in GK(56.38%).Conclusion This study specified three Sichuan local Paris resources with excellent rhizome features.And they performed well after introduced to Chengdu Plain providing the material basis for the follow-up breeding study of Paris.Three resources have medicinal potential,especially the polyphyllin Ⅰ and polyphyllin content in MH(P.forrestii) is higher,which can provide a new choice for screening substitution materials of P.polyphylla var.chinensis and P.polyphylla var.yunnanensis.
引文
[1]何俊,张舒,王红,等.滇重楼植物的研究进展[J].云南植物研究,2006,28(3):271-276.
[2]李恒.重楼属植物[M].北京:科学出版社,1998.
[3]李强,丁春邦,李燕,等.四川重楼属药用植物种类及地理分布[J].时珍国医国药,2008,19(3):629-631.
[4]李林玉,李绍平,杨斌,等.滇重楼不同居群形态变异研究[J].中国中药杂志,2010,35(22):2959-2962.
[5]Tautz D,Arctander P,Minelli A,et al.DNA points the way ahead in taxonomy[J].Nature,2002,418(6897):479-479.
[6]Ji Y H,Peter W F,Li H,et al.Phylogeny and classification of Paris(Melanthiaceae)inferred from DNA sequence data[J].Annals Bot,2006,98:245-256.
[7]中国药典[S].二部.2015.
[8]尹鸿翔,张浩,薛丹,等.川滇地区重楼属药用植物资源质量初评[J].中国中药杂志,2007,32(13):1344-1346.
[9]黄贤校,高文远,满淑丽,等.重楼属药用植物皂苷类化学成分及其生源途径的研究进展[J].中草药,2009,40(3):483-489.
[10]景松松,王颖,李雪娇,等.HPLC法测定不同产地重楼属植物中7种甾体皂苷成分[J].中草药,2012,43(12):2435-2437.
[11]李海涛,罗先文,管燕红,等.云南省不同地区滇重楼皂苷含量的对比及影响因子分析[J].中国中药杂志,2014,34(5):803-806.
[12]刘涛,赵英良,杨莹,等.滇重楼的psbA-trnH条形码分子鉴定研究[J].天然产物研究与开发,2015(27):758-762.
[13]彭世陆,刘丽芳,朱华旭,等.RP-HPLC测定中药复方癌痛平中重楼皂苷[J].中华中医药杂志,2016,31(10):4281-4283.
[14]王跃虎,牛红梅,张兆云,等.重楼属植物的药用价值及其化学物质基础[J].中国中药杂志,2015,40(5):833-839.
[15]唐玲,王艳芳,李荣英,等.滇重楼不同居群的表型多样性研究[J].中国农学通报,2013,29(18):89-95.
[16]赵俊凌,李学兰,王艳芳,等.云南省不同地区滇重楼4种皂苷含量的比较[J].中华中医药杂志,2012,27(8):2076-2078.
[17]于华会,杨志玲,杨旭,等.药用植物种质资源ITS序列研究进展[J].中草药,2010,41(3):491-496.
[18]任保青,陈之端.植物DNA条形码技术[J].植物学报,2010,45(1):1-12.
[19]陈士林,姚辉,韩建萍,等.中药材DNA条形码分子鉴定指导原则[J].中国中药杂志,2013,38(2):141-148.
[20]Zhao Y,Kang L P,Liu Y X,et al.Steroidal saponins from the rhizome of Paris polyphylla and their cytotoxic activities[J].Planta Med,2009,75(4):356-363.
[21]李朋,薛丹,文飞燕,等.长柱重楼形态描述补充及其皂苷类成分特征[J].时珍国医国药,2016,27(1):200-202.
[22]武珊珊,高文远,段宏泉,等.重楼化学成分和药理作用研究进展[J].中草药,2004,35(3):344-347.
[23]景松松,王颖,李雪娇,等.黑籽重楼化学成分及其抗肿瘤活性研究[J].中草药,2017,48(6):1093-1098.
[24]颜璐璐,张艳军,高文远,等.滇重楼皂苷成分体内外抗肺癌活性研究[J].中草药,2009,40(3):424-428.
[25]Fu Y L,Yu Z Y,Tang X M,et al.Pennogenin glycosides with aspirostanol structure are strong platelet agonists:Structural requirement for activity and mode of platelet agonist synergism[J].J Thromb Haemost,2008,6(3):524-533.
[26]Matsuda H,Pongpiriyadacha Y,Morikawa T,et al.Protectiveeffects of steroid saponins from Paris polyphylla var.yunnanensison ethanol-or indomethacininduced gastric mucosal lesions inrats:structural requirement for activity and mode of action[J].Bioorg Med Chem Lett,2003,13(6):1101-1106.
[27]李宇华,孙阳,樊磊,等.重楼皂苷VI抑制结肠癌LoVo细胞转移的作用及机制研究[J].华南国防医学杂志,2015,29(8):571-574.
[28]刘欢,何忠俊,梁社往,等.滇重楼高效液相色谱指纹图谱研究[J].中草药,2012,43(9):1846-1851.
[2]李恒.重楼属植物[M].北京:科学出版社,1998.
[3]李强,丁春邦,李燕,等.四川重楼属药用植物种类及地理分布[J].时珍国医国药,2008,19(3):629-631.
[4]李林玉,李绍平,杨斌,等.滇重楼不同居群形态变异研究[J].中国中药杂志,2010,35(22):2959-2962.
[5]Tautz D,Arctander P,Minelli A,et al.DNA points the way ahead in taxonomy[J].Nature,2002,418(6897):479-479.
[6]Ji Y H,Peter W F,Li H,et al.Phylogeny and classification of Paris(Melanthiaceae)inferred from DNA sequence data[J].Annals Bot,2006,98:245-256.
[7]中国药典[S].二部.2015.
[8]尹鸿翔,张浩,薛丹,等.川滇地区重楼属药用植物资源质量初评[J].中国中药杂志,2007,32(13):1344-1346.
[9]黄贤校,高文远,满淑丽,等.重楼属药用植物皂苷类化学成分及其生源途径的研究进展[J].中草药,2009,40(3):483-489.
[10]景松松,王颖,李雪娇,等.HPLC法测定不同产地重楼属植物中7种甾体皂苷成分[J].中草药,2012,43(12):2435-2437.
[11]李海涛,罗先文,管燕红,等.云南省不同地区滇重楼皂苷含量的对比及影响因子分析[J].中国中药杂志,2014,34(5):803-806.
[12]刘涛,赵英良,杨莹,等.滇重楼的psbA-trnH条形码分子鉴定研究[J].天然产物研究与开发,2015(27):758-762.
[13]彭世陆,刘丽芳,朱华旭,等.RP-HPLC测定中药复方癌痛平中重楼皂苷[J].中华中医药杂志,2016,31(10):4281-4283.
[14]王跃虎,牛红梅,张兆云,等.重楼属植物的药用价值及其化学物质基础[J].中国中药杂志,2015,40(5):833-839.
[15]唐玲,王艳芳,李荣英,等.滇重楼不同居群的表型多样性研究[J].中国农学通报,2013,29(18):89-95.
[16]赵俊凌,李学兰,王艳芳,等.云南省不同地区滇重楼4种皂苷含量的比较[J].中华中医药杂志,2012,27(8):2076-2078.
[17]于华会,杨志玲,杨旭,等.药用植物种质资源ITS序列研究进展[J].中草药,2010,41(3):491-496.
[18]任保青,陈之端.植物DNA条形码技术[J].植物学报,2010,45(1):1-12.
[19]陈士林,姚辉,韩建萍,等.中药材DNA条形码分子鉴定指导原则[J].中国中药杂志,2013,38(2):141-148.
[20]Zhao Y,Kang L P,Liu Y X,et al.Steroidal saponins from the rhizome of Paris polyphylla and their cytotoxic activities[J].Planta Med,2009,75(4):356-363.
[21]李朋,薛丹,文飞燕,等.长柱重楼形态描述补充及其皂苷类成分特征[J].时珍国医国药,2016,27(1):200-202.
[22]武珊珊,高文远,段宏泉,等.重楼化学成分和药理作用研究进展[J].中草药,2004,35(3):344-347.
[23]景松松,王颖,李雪娇,等.黑籽重楼化学成分及其抗肿瘤活性研究[J].中草药,2017,48(6):1093-1098.
[24]颜璐璐,张艳军,高文远,等.滇重楼皂苷成分体内外抗肺癌活性研究[J].中草药,2009,40(3):424-428.
[25]Fu Y L,Yu Z Y,Tang X M,et al.Pennogenin glycosides with aspirostanol structure are strong platelet agonists:Structural requirement for activity and mode of platelet agonist synergism[J].J Thromb Haemost,2008,6(3):524-533.
[26]Matsuda H,Pongpiriyadacha Y,Morikawa T,et al.Protectiveeffects of steroid saponins from Paris polyphylla var.yunnanensison ethanol-or indomethacininduced gastric mucosal lesions inrats:structural requirement for activity and mode of action[J].Bioorg Med Chem Lett,2003,13(6):1101-1106.
[27]李宇华,孙阳,樊磊,等.重楼皂苷VI抑制结肠癌LoVo细胞转移的作用及机制研究[J].华南国防医学杂志,2015,29(8):571-574.
[28]刘欢,何忠俊,梁社往,等.滇重楼高效液相色谱指纹图谱研究[J].中草药,2012,43(9):1846-1851.