缬草抗心律失常的药效物质及作用机理研究
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摘要
缬草(Valeriana officinalis L.)系败酱科(Valerianace)缬草属(Valerian)植物,根及根茎是主要入药部位。具有镇静安神、解痉止痛之功效,主治心神不安,心悸失眠,癫狂,风湿痹痛,痛经等。现代药理研究表明,缬草具有较好的抗心律失常作用。
为了阐释缬草抗心律失常作用的物质基础,在前人研究的基础上,采用学科交叉的研究方法,通过超临界CO_2萃取法和溶剂法,将缬草分为不同提取部位;然后以乌头碱、氯仿两种诱导剂诱导心律失常的模型为指标进行药理筛选,确定了缬草抗心律失常的主要活性部位。缬草水溶性物质部位有稳定的降低室颤发生率,推迟心律失常发生时间,减少死亡率的作用;超临界CO_2萃取物和水液部位对氯仿诱发小鼠心室纤颤具有明显的拮抗作用。氯仿、乌头碱两种诱导剂均能诱导心律失常的发生,但其诱发机理不同。缬草水溶性物质部位对两种诱导剂诱导的心律失常均有明显的对抗作用,说明其抗心律失常机制是多方面的,符合中药多成分多靶点的作用机理。
采用色谱分离法和波谱鉴定法等现代科学技术,从缬草抗心律失常有效部位中分离和鉴定了5种化合物,分别为青刺尖木脂醇苷(prinsepiol4-O-β-D-glucoside)、8-羟基松脂醇苷(8-hydroxypinoresinol-4-O-β-D-glucoside)、β-谷甾醇(β-sitosterol)、kanokoside A、绿原酸(Chlorogenic acid)。
利用现代电生理学中的膜片钳技术,将上述分离得到的化合物对转基因Kv1.5细胞进行了抗心律失常活性筛选,从中筛选出2个具有抗心律失常活性的化合物,分别为青刺尖木脂醇苷、8-羟基松脂醇苷;并对青刺尖木脂醇苷进行了作用机理探讨,研究结果表明,青刺尖木脂醇苷对Kv1.5钾通道的抑制,说明青刺尖木脂醇苷的抗心律失常可能主要是通过抑制钾通道,延长动作电位和有效不应期而达到治疗目的,初步阐释了缬草抗心律失常的作用机制,所以青刺尖木脂醇苷有望成为治疗抗心律失常药。
在抗心律失常物质基础研究的基础上,采用高效液相色谱法对缬草水溶性部位进行了指纹图谱研究,标定了15个共有峰,归属了4个共有峰,分别是青刺尖木脂醇苷、8-羟基松脂醇苷、绿原酸和橙皮苷;并对不同产地、不同采收时间以及缬草近缘品种进行了比较;建立了可靠的分析方法,符合《国家食品药品监督管理局》颁布的中药注射剂指纹图谱技术要求。从而为规范缬草属植物的药用来源及采收季节提供了参考依据。
采用高效液相色谱法对缬草中抗心律失常有效成分青刺尖木脂醇苷进行了含量分析研究,建立了灵敏度高、重现性好、可靠性强的含量测定方法。从而为有效控制缬草药材的质量,保证用药安全提供了科学依据。
Valeriana officinalis L.belongs to department of Patrinia of valerian,its roots and rhizomes are the main parts of medicine which have sedative and spasmolytic effects, attending uneasy,palpitation insomnia,mania,rheumatism Bitongkang,dysmenorrheal,etc.Modern pharmacological studies have shown that valerian has anti-arrhythmic effect.
In order to clarify the material basis of the anti-arrhythmic effects of Valeriana we adopted interdisciplinary research methods based on the previous studies.Through supercritical CO_2 extraction method and solvent method the Valeriana was divided into different valerian extracts,and then using aconitine alkali and chloroform to induce arrhythmia model for pharmacological screening indicators to determine the main active anti-arrhythmic site of Valeriana.The water-soluble parts of Valerian stably decrease the incidence of ventricular fibrillation,postpone arrhythmias time and reduce the mortality.Supercritical CO_2 extract and water-liquid parts have more pronounced antagonism to chloroform-induced ventricular fibrillation in mice.And the other parts also have anti-arrhythmic effects indifferent extent.Although inducing arrhythmia through different mechanisms,the water-soluble part of Valerian have altogether obvious antagonistic effects to the two agent chloroform and aconitine induced arrhythmia, while the other parts have different response.The all above reflect the mechanisms for its anti-arrhythmic is multifaceted and there are multiple active ingredients.
Using chromatographic separation and spectral identification method,we can separate and identify five kinds of compounds from the effective anti-arrhythmic parts of Valeriana as follows:Green barbed tip lignan glycosides (prinsepiol-4-0-β-D-glucoside);8-hydroxy resin glycosides (8-hydroxypinoresinol -4-o-β-D-glucoside);β-sitosterol; kanokoside A;Chlorogenic acid.
Using modern electrophysiological patch-clamp technique to investigate these compounds separated above,there are two compounds that have been proved had anti-arrhythmic activity: Green barbed tip lignan glycosides and 8-hydroxy resin glycosides.We explored the action mechanism of the green lignan glycosides and the results show that the Green barbed tip lignan glycosides through inhibiting potassium channels and prolonging effective refractory period to achieve the anti-arrhythmic therapeutic purposes.Green barbed tip lignan glycosides are expected to become anti-arrhythmic drug.The paper initially explain the mechanism of its anti-arrhythmic. On the basis of basic research of anti-arrhythmic substances we utilized high-performance liquid chromatography to research the fingerprint of water-soluble parts of Valeriana and demarcate 15 common peaks.Valeriana from different original Sources,different harvesting time and Valeriana close related species were also compared,establishing a reliable analytical method in accordance with the "Chinese Pharmacopoeia" and the State Food and Drug Administration issued technical standards.Thereby the research provides a reference for the Valeriana medicinal plants harvesting sources and season.
Using high-performance liquid chromatography to analyze active anti-arrhythmic ingredients Green barbed tip lignan glycosides of Valeriana content has high sensitivity,good reproducibility,powerful reliability,through which we can effectively control Valeriana medicinal quality and provide a scientific basis to ensure drug safety.
为了阐释缬草抗心律失常作用的物质基础,在前人研究的基础上,采用学科交叉的研究方法,通过超临界CO_2萃取法和溶剂法,将缬草分为不同提取部位;然后以乌头碱、氯仿两种诱导剂诱导心律失常的模型为指标进行药理筛选,确定了缬草抗心律失常的主要活性部位。缬草水溶性物质部位有稳定的降低室颤发生率,推迟心律失常发生时间,减少死亡率的作用;超临界CO_2萃取物和水液部位对氯仿诱发小鼠心室纤颤具有明显的拮抗作用。氯仿、乌头碱两种诱导剂均能诱导心律失常的发生,但其诱发机理不同。缬草水溶性物质部位对两种诱导剂诱导的心律失常均有明显的对抗作用,说明其抗心律失常机制是多方面的,符合中药多成分多靶点的作用机理。
采用色谱分离法和波谱鉴定法等现代科学技术,从缬草抗心律失常有效部位中分离和鉴定了5种化合物,分别为青刺尖木脂醇苷(prinsepiol4-O-β-D-glucoside)、8-羟基松脂醇苷(8-hydroxypinoresinol-4-O-β-D-glucoside)、β-谷甾醇(β-sitosterol)、kanokoside A、绿原酸(Chlorogenic acid)。
利用现代电生理学中的膜片钳技术,将上述分离得到的化合物对转基因Kv1.5细胞进行了抗心律失常活性筛选,从中筛选出2个具有抗心律失常活性的化合物,分别为青刺尖木脂醇苷、8-羟基松脂醇苷;并对青刺尖木脂醇苷进行了作用机理探讨,研究结果表明,青刺尖木脂醇苷对Kv1.5钾通道的抑制,说明青刺尖木脂醇苷的抗心律失常可能主要是通过抑制钾通道,延长动作电位和有效不应期而达到治疗目的,初步阐释了缬草抗心律失常的作用机制,所以青刺尖木脂醇苷有望成为治疗抗心律失常药。
在抗心律失常物质基础研究的基础上,采用高效液相色谱法对缬草水溶性部位进行了指纹图谱研究,标定了15个共有峰,归属了4个共有峰,分别是青刺尖木脂醇苷、8-羟基松脂醇苷、绿原酸和橙皮苷;并对不同产地、不同采收时间以及缬草近缘品种进行了比较;建立了可靠的分析方法,符合《国家食品药品监督管理局》颁布的中药注射剂指纹图谱技术要求。从而为规范缬草属植物的药用来源及采收季节提供了参考依据。
采用高效液相色谱法对缬草中抗心律失常有效成分青刺尖木脂醇苷进行了含量分析研究,建立了灵敏度高、重现性好、可靠性强的含量测定方法。从而为有效控制缬草药材的质量,保证用药安全提供了科学依据。
Valeriana officinalis L.belongs to department of Patrinia of valerian,its roots and rhizomes are the main parts of medicine which have sedative and spasmolytic effects, attending uneasy,palpitation insomnia,mania,rheumatism Bitongkang,dysmenorrheal,etc.Modern pharmacological studies have shown that valerian has anti-arrhythmic effect.
In order to clarify the material basis of the anti-arrhythmic effects of Valeriana we adopted interdisciplinary research methods based on the previous studies.Through supercritical CO_2 extraction method and solvent method the Valeriana was divided into different valerian extracts,and then using aconitine alkali and chloroform to induce arrhythmia model for pharmacological screening indicators to determine the main active anti-arrhythmic site of Valeriana.The water-soluble parts of Valerian stably decrease the incidence of ventricular fibrillation,postpone arrhythmias time and reduce the mortality.Supercritical CO_2 extract and water-liquid parts have more pronounced antagonism to chloroform-induced ventricular fibrillation in mice.And the other parts also have anti-arrhythmic effects indifferent extent.Although inducing arrhythmia through different mechanisms,the water-soluble part of Valerian have altogether obvious antagonistic effects to the two agent chloroform and aconitine induced arrhythmia, while the other parts have different response.The all above reflect the mechanisms for its anti-arrhythmic is multifaceted and there are multiple active ingredients.
Using chromatographic separation and spectral identification method,we can separate and identify five kinds of compounds from the effective anti-arrhythmic parts of Valeriana as follows:Green barbed tip lignan glycosides (prinsepiol-4-0-β-D-glucoside);8-hydroxy resin glycosides (8-hydroxypinoresinol -4-o-β-D-glucoside);β-sitosterol; kanokoside A;Chlorogenic acid.
Using modern electrophysiological patch-clamp technique to investigate these compounds separated above,there are two compounds that have been proved had anti-arrhythmic activity: Green barbed tip lignan glycosides and 8-hydroxy resin glycosides.We explored the action mechanism of the green lignan glycosides and the results show that the Green barbed tip lignan glycosides through inhibiting potassium channels and prolonging effective refractory period to achieve the anti-arrhythmic therapeutic purposes.Green barbed tip lignan glycosides are expected to become anti-arrhythmic drug.The paper initially explain the mechanism of its anti-arrhythmic. On the basis of basic research of anti-arrhythmic substances we utilized high-performance liquid chromatography to research the fingerprint of water-soluble parts of Valeriana and demarcate 15 common peaks.Valeriana from different original Sources,different harvesting time and Valeriana close related species were also compared,establishing a reliable analytical method in accordance with the "Chinese Pharmacopoeia" and the State Food and Drug Administration issued technical standards.Thereby the research provides a reference for the Valeriana medicinal plants harvesting sources and season.
Using high-performance liquid chromatography to analyze active anti-arrhythmic ingredients Green barbed tip lignan glycosides of Valeriana content has high sensitivity,good reproducibility,powerful reliability,through which we can effectively control Valeriana medicinal quality and provide a scientific basis to ensure drug safety.
引文
[1]陈虎彪,朱建民,赵玉英等.药用缬草及其中国产近缘种的化学分类学研究.中成药,1999;21(12):652-654
[2]李时珍.《本草纲目》(校对本)第2册,北京:人民卫生出版社,1979,845.
[3]中华人民共和国卫生部药典委员会编.中华人民共和国药典.1977年版,一部,北京:人民卫生出版社,1978,633.
[4]石晋丽.国产缬草属药用植物资源的研究.北京中医药大学博士学位论文,2004.
[5]黄宝康.中国缬草属植物的生药学及缬草的种内变异研究.第二军医大学博士学位论文,2005.
[6]黄宝康,郑汉臣,秦路平,等.国产缬草属药用植物资源调查.中药材,2004;27(9):632-634.
[7]陈磊.中国缬草属药用植物的生药鉴定和资源利用研究.第二军医大学硕士学位论文,2002.
[8]兰茂著,《滇南本草》整理组整理.滇南本草.第2卷,昆明:云南人民出版社,1977,273.
[9]郭济贤.中国缬草属植物叶毛茸的显微观察.上海第一医学院学报,1985:12(4):260-262.
[10]江泽荣.毛节缬草的生药学研究.药学学报,1964;11(11):778.
[11]明东升.中药蜘蛛香和缬草类的形态组织学研究.上海医科大学学报,1993;20(3):205-207.
[12]石晋丽,刘勇,肖培根.缬草属植物化学成分与药理作用.国外医药·植物药分册,2003;18(6):231-239.
[13]Picinell A L,arena S new lignans from the roots of Valeriana prionophylla with an antioxdative and vasorelaxant activities.Nat Prod,2004;67(7):1135-1140.
[14]Britta Schumacher,Silke Scholle,Josef Holzl,et al.Lignans Isolated from Valerian:Identification and Characterization of a New Olivil Derivative with Partial Agonistic Activity at Al Adenosine Receptors.Nat.Prod,2002;65:1479-1485.
[15]Femandez S,WasowskiC.sedative and sleep-enhancing properties of linarin,a flavonoid-isolated from Valeriana officinalis.Phamaool B iochen B ehav,2004;77(2):399-404.
[16]徐红,袁惠南,潘丽华,等.缬草挥发油对中枢神经系统药理作用研究.药物分析杂志,1997;17(6):399-401.
[17]hiller K O.Phytother Res.1996;10(2):145-151.
[18]Malva J O.Neuroprotective properties of Valeriana officinalis extrats.Neurotox Res,2004;6(2):131-140.
[19]吉富史郎.Natural Medicines.2 000;54(2):55-60.
[20]曹斌,洪庚辛,等.蜘蛛香的中枢抑制作用.中国中药杂志,1994;19(1):40-42.
[21]Oshima Y.Chem Pharm Bull.1995;43(1):169-170.
[22]张宝恒,孟和平,王彤,等.缬草乙醇提取物(VES)对心血管的作用.药学学报,1982;17(5):382.
[23]杨晶,薛存宽,朱咸中,等.86Rb摄取示踪法评定几种中药提取物对心、肾组织微循环灌流量的影响.微循环学杂志,1998;8(1):15-17.
[24]杨晶.神农缬草挥发油的平喘作用.中西医结合杂志,1988;8:28.
[25]陈灼芳.墨香挥发油抗心律失常作用的研究.中西医结合杂志,1990:10(10):614.
[26]王玉良,徐森根,范世藩.缬草抗心律失常的心肌细胞电生理学观察.中华心血管病杂志,1979;7(4):275-282.
[27]袁韶华,张宝恒,吴凤简.缬草提取物(V3d)抗心律失常的实验研究.中药药理与临床,1989;5(6):15-18.
[28]贾健宁,张宝恒.缬草提取物(V3d)对心血管系统的作用.广西中医学 院学报,1999;16(1):40-42.
[29]黄峥嵘.缬草提取物对兔心室肌细胞离子通道的影响.武汉大学硕士学位论文.2004.5:22.
[30]薛存宽,杨静,等.缬草提取物药效学研究与工艺探讨.中药新药与临床药理,1996;7(3):15-19.
[31]胡昌兴.宽叶缬草对高脂血症家兔血脂代谢的影响.海军医高专学报,1999;21(2):65-68.
[32]Keochanthal a-Bounthanh C.Phytotherapy Resear ch.1993;7:124-127.
[33]Bos R.Phytomedicine.1998;5(3):219-225.
[34]Fursa N S.Farmatsiva(Moscow).1993;42(6):76.
[35]陈玲,贾汝汉,丁国华,等.缬草油对2型糖尿病大鼠肾脏的保护作用及其机制探讨.中华肾脏病杂志,2003;19(3):168-172.
[36]唐久余,曾园山,陈巧格,等.缬草对慢性应激导致的抑郁大鼠大脑海马5-轻色胺水平、细胞增殖及神经元数量的影响.中西医结合学报,2008;6(3):283-288.
[37]吴波,付玉梅,黄爱华,等.缬草挥发油对戊四氮致癫痫大鼠海马内γ-氨基丁酸和谷氨酸含量的影响.中华中医药学刊,2008;26(11):2476-2477.
[38]陈素美,杨荣时,曾玉兰,等.缬草提取物对大鼠实验性肺纤维化的干预作用.医药导报,2008;27(11):1295-1297.
[39]Kastke,Floyd A(USA).U.S pat Appl.Publ.US 20010025059A127 Sep 2001;9pp.;Cont-in-part of U S.Ser.No.175,286(English).
[40]Muraveva D A.Farmatsiya(Moscow),1994;43(3):62.
[41]Fursa N S.Farmatsiya(Moscow),1993;43(1):69.
[42]Aleksandrov S E.Farmatsiya(Moscow),1994;43(2):65.
[43]Bolshakowa,I.V.Biofizika(Russian),1997;42(2):480-483.
[44]Inatle,D J.Et hnophar rracol,2000;72(1,2):47-51.
[45]Mizumoto,Daigo.Jpn.Kokai Tokkyo Koho JP 2001131026 A2 15May,2001;10pp.(Japan).
[46]詹国瑛,孙建琴,孙晓红.缬草的毒性及致突变性测试.癌变·畸变·突变,1996;8(3):180.
[47]Tufik Sergio.Journal of thnopharmacology,1994;41(1-2):39.
[48]Monograph Valeriana officinalis.Alfem Med Rev,2004;9(4):438-441.
[49]Kohnen R,Oswald W D.The effects of Valerian,propranolol and their combination oactivation,performance,and mood of healthy volunteers under social stress conditions.Pharmcopsychiatry,1988;21:447-448.
[50]Eadie M J Could Valerian have been the first anticonvul sant.Epilepsia,2004;45(11):1338-1343.
[51]Herbalgram.1996;(36):16.
[52]都晓伟,吴军凯.缬草属植物化学成分及药理活性研究进展.国外医药·植物药分册,2006;21(1):10-14.
[53]杨桂元,王玮.宽叶缬草治疗冠心病的临床研究.中国中西医结合杂志,1994;14(9):540-542.
[54]黄峥嵘,唐其柱,史锡腾,等.非缬草单萜氧化物对单个兔心室肌细胞钠通道的影响.中国心脏起搏与心电生理杂志,2004;18(3):214-217.
[55]黄峥嵘,唐其柱,张丽娟等.缬草单萜氧化物对兔心室肌线粒体ATP 敏感性钾通道的影响.中国中医与微循环杂志,2005;15(1):69-71.
[56]徐淑云,卞如濂,陈修主编.药理实验方法,第3版.北京:人民卫生出版社,2003,1172.
[57]汪晖,吴基良.药理学实验,武汉:湖北科技出版社,2002,63.
[58]张建超,方颖,刘焱文,等正交法优化缬草挥发油超临界CO_2流体提取工艺.中成药,2008;30(2):201-203.
[59]王辉,罗顺德.二乙酰基莲心碱抗实验性心律失常的作用.中国医院药学杂志,2001;21(6):328.
[60]龚冬梅,单宏丽,周宇宏,等.哇巴因和乌头碱诱发豚鼠和大鼠心律失常的离子作用靶点.药学学报,2004;39(5):328-332.
[61]马亮,杨娇,傅善权,等.野生与家种缬草挥发油GC-MS分析.重庆工学院学报(自然科学版),2007;21(5):119-123.
[62]吴筑平,刘密新,姚焕新,等.缬草挥发油化学成分的研究.中国中药学杂志,1999;34(11):733-735.
[63]陈虎彪,徐学明,赵玉英.毛节缬草挥发油化学成分的研究.中国中药杂志,1998;23(7):418-421.
[64]王立群,熊义涛,陶福华,等.宽叶缬草挥发油成分分析.中药材,1999:22(6):298-301.
[65]周琳,张悦.野生黑水缬草和北缬草根精油的分析.国土与自然资源研究,2006;2:92-93.
[66]Safaralie A,Fatam iS,Sefidkon F.E ssential oil composition of Valeria and officinalis L.roots cultivated in Iran comparative analysis between supercritical CO_2 extraction and hydrod istillation.J Chrum atogrA,2008;1180(1-2):159-162.
[67]明东升,郭济贤,顺庆生,等.四种缬草生药挥发油化学成分的气相层析一质谱联用鉴定.中成药,1994;16(1):41-43.
[68]谷臣华,吕盛槐,谷力.缬草花油的提取与GC-MS测定.林产化学与工业,1999:19(3):75-78.
[69]周霆,黄宝康.缬草挥发油的化学成分及药理活性研究进展.时珍国医国药,2008;19(11):2663-2664.
[70]Anna Lisa Piccinelli,Swizly Arana,Armando Caceres,et.New Lignans from the Roots of Valeriana prionophylla with Antioxidative and Vasorelaxant Activities.Nat.Prod.2004;67:1135-1140.
[71]Britta Schumacher,Silke Scholle,Josef Ho··lzl,Lignans Isolated from Valerian:Identification and Characterization of a New Olivil Derivative with Partial Agonistic Activity at A1Adenosine Receptors.Nat.Prod.2002;65:479-1485.
[72]Sun LN,Hong YF.Chemical Constituents of Chaenomeles sinensis (Thouin.) Koehne.China Pharm,2000;9(1):6-9.
[73]Ays,e Kuruu··zu··m-Uz,Zuhal Gu··venalp,L.O··mu··r Demirezer,et.40-Deoxy iridoid glycosides from Centranthus longiflorus.Phytochemistry 2002;6(1):937-941.
[74]张赞彬,谈毅婷.甘薯渣中多酚类化合物的分离与结构鉴定.食品研究与开发,2007;28(12):120-122.
[75]Hamill OP,Many A,Neher E,Sakmann B,Siqworth FJ.improvedpatch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.Pf lugers Arch,1981;391(2):85-100.
[76]xiao Gs,zhou JJ,Wang Gy,et al.In vitro Electrophy siologic effects of morphine in Rabbit.Ventricular myocytes Anesthesiology,2005;103(2):280-286.
[77]李泱.离子通道与膜片钳技术.甘肃:甘肃文化出版社,2001,151
[78]杜景霞,张晓华,胡燕,等.白黎芦醇对豚鼠心室肌细胞钠电流的影响.中国现代应用药学杂志,2008;25(4):290-293.
[79]王赫,周宇宏,单宏丽,等.冬虫夏草水提液对豚鼠及人鼠心室肌细胞钾通道的作用.中国药理学通报,2004;20(5):536-539.
[80]张绪国,王家宁,李建军,等.银杏叶提取物对豚鼠心室肌细胞动作电位及L型钙离子通道的影响.中国临床药理学与治疗学,2004;9(3):345-349.
[81]Deng C Y,Lin S G,Yang M,et al.Effect of carvedilol on sodiun channel current in isolated ventricular myocytes of rats.Chin Phamacol Bull,2005;21(5):568-572.
[82]邝素娟,邓春玉,钱卫民,等.醋柳黄酮对大鼠心室肌细胞瞬间外向整流钾电流的影响.国药理学通报,2007;23(9):1237-1240.
[83]聂宏光,陈磊,王岩,等.羧甲基壳聚糖对大鼠单一心室肌细胞ATP 敏感钾电流的作用.中国海洋药物杂志,2008;27(1):26-27.
[84]邓春玉,唐承静,邝素娟,等.灯盏花素对大鼠心室肌细胞瞬间外向和内向整流钾电流的影响.中国病理生理杂志,2008;24(1):84-88.
[85]李红,张爽,石卓,等.蒺藜皂苷对成年豚鼠心室肌细胞钾电流的影响.吉林大学学报(医学版),2008;34(6):927-930.
[86]张华.内向整流钾通道—葛根素防治心律失常及其机制的实验研究.第四军医大学博士学位论文;2002.5.
[87]李晓光,张文杰,孙莹,等.黄芪总黄酮对豚鼠心室肌细胞动作电位和延迟整流钾电流的作用.心肺血管病杂志,2008;27(1):49-51.
[88]孙宏丽,董德利,李磊,等.三氧化二砷对豚鼠心室肌细胞钾电流的影响.中国药学杂志,2008;43(24)1867-1870.
[89]高分飞,黄展勤,周燕琼,等.心肌细胞分离方法的改进及其钾电流的观察.广西医科大学学报,2008;25(3)338-340.
[90]陈修.心血管药理学[M].北京:人民卫生出版社,1989:483-486.
[91]Dorian P.Antiarrhythmic drug therapy of atrial fibrillation:focus on new agents.Cardiovasc Pharmacol,2003;8:S27-S31
[92]Peukert S,Brendel J,Pirard B,et al.Identification,synthesis and activity of novel blockers of the voltagegated potassium channel Kv1.5.J Med Chem,2003;46:486-498.
[93]Noda Y,Anzai K,Mori A,et al.Shirunei M and PackerL.Hyclroxyl and superoxide anion radical scavenging activitiesof natural source antioxidants using the computerized JES-FR30ESR spectrometer system.Biochem Mol Biol Int,1997;42(1):35-44.
[92]Pietri S,Maureli E,Drieu K,et al.Cardioprotective andantioxidant effects of the terpenoid constituents of ginkgo biloba extract(EGB761).J MolCell Cardio,1997;29(2):733-742.
[95]国家中医药管理局中华本草编委会.中华本草.第7期.第12卷,上海:上海科技出版社,1999,574.
[96]周玉新.中药指纹图谱研究技术,北京:化学工业出版社,2002:108.
[97]陈磊,康鲁平,秦路平,等.缬草属药用植物HPLC指纹特征研究.中药材,2003;26(4):256-259.
[98]石晋丽,刘勇,肖培根.缬草属药用植物极性成分的HPLC指纹图谱研究.中国中药杂志,2005;30(6):427-429.
[99]《中药注射剂指纹图谱研究的技术要求(暂行)的通知》.中国药品标准,2000;1(4):3.
[100]崔亚君,穆赫塔尔·亚森,岳松健,等.七种缬草属植物中缬草三酯和乙酰缬草三酯含量考察.西北药学杂志,1999;14(4):152-153.
[101]黄仁泉,张立,杨建丽.不同种缬草中缬草三酯和缬草烯酸类成分的HPLC的分析和比较.中草药,2002,33(11):1000-1001.
[102]傅亮,楚清脆,黄宝康,等.毛细管电泳一电化学检测法测定蜘蛛香中多元酚类化合物.分析化学,2005;33(2):161-164.
[103]明东升,郭济贤.缬草类生药中总缬草素的含量测定.上海医科大学学报,1993;20(3):210-212.
[104]陈磊,康鲁平,秦路平,等.总缬草素的质量标准和镇静催眠活性研究.中成药,2003;25(8):663-665.
[105]陈磊,秦路平,郑汉臣,等.三种缬草属植物的缬草素类含量种间和种内比较.中药材,2002;25(4):237-238.
[106]张素霞.缬草根和根茎中的挥发油、缬草酸及其衍生物和缬草三酯含量的季节变化及药用植株选择.国外医药.植物药分册,1999;14(1):21-22.
[107]左月明.黑水缬草抗老年痴呆有效部位化学成分和药理作用研究.黑龙江中医药大学博士学位论文,2007,5.
[108]中国药典.2005年版,一部,北京:人民卫生出版社,2005:附录33.
[2]李时珍.《本草纲目》(校对本)第2册,北京:人民卫生出版社,1979,845.
[3]中华人民共和国卫生部药典委员会编.中华人民共和国药典.1977年版,一部,北京:人民卫生出版社,1978,633.
[4]石晋丽.国产缬草属药用植物资源的研究.北京中医药大学博士学位论文,2004.
[5]黄宝康.中国缬草属植物的生药学及缬草的种内变异研究.第二军医大学博士学位论文,2005.
[6]黄宝康,郑汉臣,秦路平,等.国产缬草属药用植物资源调查.中药材,2004;27(9):632-634.
[7]陈磊.中国缬草属药用植物的生药鉴定和资源利用研究.第二军医大学硕士学位论文,2002.
[8]兰茂著,《滇南本草》整理组整理.滇南本草.第2卷,昆明:云南人民出版社,1977,273.
[9]郭济贤.中国缬草属植物叶毛茸的显微观察.上海第一医学院学报,1985:12(4):260-262.
[10]江泽荣.毛节缬草的生药学研究.药学学报,1964;11(11):778.
[11]明东升.中药蜘蛛香和缬草类的形态组织学研究.上海医科大学学报,1993;20(3):205-207.
[12]石晋丽,刘勇,肖培根.缬草属植物化学成分与药理作用.国外医药·植物药分册,2003;18(6):231-239.
[13]Picinell A L,arena S new lignans from the roots of Valeriana prionophylla with an antioxdative and vasorelaxant activities.Nat Prod,2004;67(7):1135-1140.
[14]Britta Schumacher,Silke Scholle,Josef Holzl,et al.Lignans Isolated from Valerian:Identification and Characterization of a New Olivil Derivative with Partial Agonistic Activity at Al Adenosine Receptors.Nat.Prod,2002;65:1479-1485.
[15]Femandez S,WasowskiC.sedative and sleep-enhancing properties of linarin,a flavonoid-isolated from Valeriana officinalis.Phamaool B iochen B ehav,2004;77(2):399-404.
[16]徐红,袁惠南,潘丽华,等.缬草挥发油对中枢神经系统药理作用研究.药物分析杂志,1997;17(6):399-401.
[17]hiller K O.Phytother Res.1996;10(2):145-151.
[18]Malva J O.Neuroprotective properties of Valeriana officinalis extrats.Neurotox Res,2004;6(2):131-140.
[19]吉富史郎.Natural Medicines.2 000;54(2):55-60.
[20]曹斌,洪庚辛,等.蜘蛛香的中枢抑制作用.中国中药杂志,1994;19(1):40-42.
[21]Oshima Y.Chem Pharm Bull.1995;43(1):169-170.
[22]张宝恒,孟和平,王彤,等.缬草乙醇提取物(VES)对心血管的作用.药学学报,1982;17(5):382.
[23]杨晶,薛存宽,朱咸中,等.86Rb摄取示踪法评定几种中药提取物对心、肾组织微循环灌流量的影响.微循环学杂志,1998;8(1):15-17.
[24]杨晶.神农缬草挥发油的平喘作用.中西医结合杂志,1988;8:28.
[25]陈灼芳.墨香挥发油抗心律失常作用的研究.中西医结合杂志,1990:10(10):614.
[26]王玉良,徐森根,范世藩.缬草抗心律失常的心肌细胞电生理学观察.中华心血管病杂志,1979;7(4):275-282.
[27]袁韶华,张宝恒,吴凤简.缬草提取物(V3d)抗心律失常的实验研究.中药药理与临床,1989;5(6):15-18.
[28]贾健宁,张宝恒.缬草提取物(V3d)对心血管系统的作用.广西中医学 院学报,1999;16(1):40-42.
[29]黄峥嵘.缬草提取物对兔心室肌细胞离子通道的影响.武汉大学硕士学位论文.2004.5:22.
[30]薛存宽,杨静,等.缬草提取物药效学研究与工艺探讨.中药新药与临床药理,1996;7(3):15-19.
[31]胡昌兴.宽叶缬草对高脂血症家兔血脂代谢的影响.海军医高专学报,1999;21(2):65-68.
[32]Keochanthal a-Bounthanh C.Phytotherapy Resear ch.1993;7:124-127.
[33]Bos R.Phytomedicine.1998;5(3):219-225.
[34]Fursa N S.Farmatsiva(Moscow).1993;42(6):76.
[35]陈玲,贾汝汉,丁国华,等.缬草油对2型糖尿病大鼠肾脏的保护作用及其机制探讨.中华肾脏病杂志,2003;19(3):168-172.
[36]唐久余,曾园山,陈巧格,等.缬草对慢性应激导致的抑郁大鼠大脑海马5-轻色胺水平、细胞增殖及神经元数量的影响.中西医结合学报,2008;6(3):283-288.
[37]吴波,付玉梅,黄爱华,等.缬草挥发油对戊四氮致癫痫大鼠海马内γ-氨基丁酸和谷氨酸含量的影响.中华中医药学刊,2008;26(11):2476-2477.
[38]陈素美,杨荣时,曾玉兰,等.缬草提取物对大鼠实验性肺纤维化的干预作用.医药导报,2008;27(11):1295-1297.
[39]Kastke,Floyd A(USA).U.S pat Appl.Publ.US 20010025059A127 Sep 2001;9pp.;Cont-in-part of U S.Ser.No.175,286(English).
[40]Muraveva D A.Farmatsiya(Moscow),1994;43(3):62.
[41]Fursa N S.Farmatsiya(Moscow),1993;43(1):69.
[42]Aleksandrov S E.Farmatsiya(Moscow),1994;43(2):65.
[43]Bolshakowa,I.V.Biofizika(Russian),1997;42(2):480-483.
[44]Inatle,D J.Et hnophar rracol,2000;72(1,2):47-51.
[45]Mizumoto,Daigo.Jpn.Kokai Tokkyo Koho JP 2001131026 A2 15May,2001;10pp.(Japan).
[46]詹国瑛,孙建琴,孙晓红.缬草的毒性及致突变性测试.癌变·畸变·突变,1996;8(3):180.
[47]Tufik Sergio.Journal of thnopharmacology,1994;41(1-2):39.
[48]Monograph Valeriana officinalis.Alfem Med Rev,2004;9(4):438-441.
[49]Kohnen R,Oswald W D.The effects of Valerian,propranolol and their combination oactivation,performance,and mood of healthy volunteers under social stress conditions.Pharmcopsychiatry,1988;21:447-448.
[50]Eadie M J Could Valerian have been the first anticonvul sant.Epilepsia,2004;45(11):1338-1343.
[51]Herbalgram.1996;(36):16.
[52]都晓伟,吴军凯.缬草属植物化学成分及药理活性研究进展.国外医药·植物药分册,2006;21(1):10-14.
[53]杨桂元,王玮.宽叶缬草治疗冠心病的临床研究.中国中西医结合杂志,1994;14(9):540-542.
[54]黄峥嵘,唐其柱,史锡腾,等.非缬草单萜氧化物对单个兔心室肌细胞钠通道的影响.中国心脏起搏与心电生理杂志,2004;18(3):214-217.
[55]黄峥嵘,唐其柱,张丽娟等.缬草单萜氧化物对兔心室肌线粒体ATP 敏感性钾通道的影响.中国中医与微循环杂志,2005;15(1):69-71.
[56]徐淑云,卞如濂,陈修主编.药理实验方法,第3版.北京:人民卫生出版社,2003,1172.
[57]汪晖,吴基良.药理学实验,武汉:湖北科技出版社,2002,63.
[58]张建超,方颖,刘焱文,等正交法优化缬草挥发油超临界CO_2流体提取工艺.中成药,2008;30(2):201-203.
[59]王辉,罗顺德.二乙酰基莲心碱抗实验性心律失常的作用.中国医院药学杂志,2001;21(6):328.
[60]龚冬梅,单宏丽,周宇宏,等.哇巴因和乌头碱诱发豚鼠和大鼠心律失常的离子作用靶点.药学学报,2004;39(5):328-332.
[61]马亮,杨娇,傅善权,等.野生与家种缬草挥发油GC-MS分析.重庆工学院学报(自然科学版),2007;21(5):119-123.
[62]吴筑平,刘密新,姚焕新,等.缬草挥发油化学成分的研究.中国中药学杂志,1999;34(11):733-735.
[63]陈虎彪,徐学明,赵玉英.毛节缬草挥发油化学成分的研究.中国中药杂志,1998;23(7):418-421.
[64]王立群,熊义涛,陶福华,等.宽叶缬草挥发油成分分析.中药材,1999:22(6):298-301.
[65]周琳,张悦.野生黑水缬草和北缬草根精油的分析.国土与自然资源研究,2006;2:92-93.
[66]Safaralie A,Fatam iS,Sefidkon F.E ssential oil composition of Valeria and officinalis L.roots cultivated in Iran comparative analysis between supercritical CO_2 extraction and hydrod istillation.J Chrum atogrA,2008;1180(1-2):159-162.
[67]明东升,郭济贤,顺庆生,等.四种缬草生药挥发油化学成分的气相层析一质谱联用鉴定.中成药,1994;16(1):41-43.
[68]谷臣华,吕盛槐,谷力.缬草花油的提取与GC-MS测定.林产化学与工业,1999:19(3):75-78.
[69]周霆,黄宝康.缬草挥发油的化学成分及药理活性研究进展.时珍国医国药,2008;19(11):2663-2664.
[70]Anna Lisa Piccinelli,Swizly Arana,Armando Caceres,et.New Lignans from the Roots of Valeriana prionophylla with Antioxidative and Vasorelaxant Activities.Nat.Prod.2004;67:1135-1140.
[71]Britta Schumacher,Silke Scholle,Josef Ho··lzl,Lignans Isolated from Valerian:Identification and Characterization of a New Olivil Derivative with Partial Agonistic Activity at A1Adenosine Receptors.Nat.Prod.2002;65:479-1485.
[72]Sun LN,Hong YF.Chemical Constituents of Chaenomeles sinensis (Thouin.) Koehne.China Pharm,2000;9(1):6-9.
[73]Ays,e Kuruu··zu··m-Uz,Zuhal Gu··venalp,L.O··mu··r Demirezer,et.40-Deoxy iridoid glycosides from Centranthus longiflorus.Phytochemistry 2002;6(1):937-941.
[74]张赞彬,谈毅婷.甘薯渣中多酚类化合物的分离与结构鉴定.食品研究与开发,2007;28(12):120-122.
[75]Hamill OP,Many A,Neher E,Sakmann B,Siqworth FJ.improvedpatch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.Pf lugers Arch,1981;391(2):85-100.
[76]xiao Gs,zhou JJ,Wang Gy,et al.In vitro Electrophy siologic effects of morphine in Rabbit.Ventricular myocytes Anesthesiology,2005;103(2):280-286.
[77]李泱.离子通道与膜片钳技术.甘肃:甘肃文化出版社,2001,151
[78]杜景霞,张晓华,胡燕,等.白黎芦醇对豚鼠心室肌细胞钠电流的影响.中国现代应用药学杂志,2008;25(4):290-293.
[79]王赫,周宇宏,单宏丽,等.冬虫夏草水提液对豚鼠及人鼠心室肌细胞钾通道的作用.中国药理学通报,2004;20(5):536-539.
[80]张绪国,王家宁,李建军,等.银杏叶提取物对豚鼠心室肌细胞动作电位及L型钙离子通道的影响.中国临床药理学与治疗学,2004;9(3):345-349.
[81]Deng C Y,Lin S G,Yang M,et al.Effect of carvedilol on sodiun channel current in isolated ventricular myocytes of rats.Chin Phamacol Bull,2005;21(5):568-572.
[82]邝素娟,邓春玉,钱卫民,等.醋柳黄酮对大鼠心室肌细胞瞬间外向整流钾电流的影响.国药理学通报,2007;23(9):1237-1240.
[83]聂宏光,陈磊,王岩,等.羧甲基壳聚糖对大鼠单一心室肌细胞ATP 敏感钾电流的作用.中国海洋药物杂志,2008;27(1):26-27.
[84]邓春玉,唐承静,邝素娟,等.灯盏花素对大鼠心室肌细胞瞬间外向和内向整流钾电流的影响.中国病理生理杂志,2008;24(1):84-88.
[85]李红,张爽,石卓,等.蒺藜皂苷对成年豚鼠心室肌细胞钾电流的影响.吉林大学学报(医学版),2008;34(6):927-930.
[86]张华.内向整流钾通道—葛根素防治心律失常及其机制的实验研究.第四军医大学博士学位论文;2002.5.
[87]李晓光,张文杰,孙莹,等.黄芪总黄酮对豚鼠心室肌细胞动作电位和延迟整流钾电流的作用.心肺血管病杂志,2008;27(1):49-51.
[88]孙宏丽,董德利,李磊,等.三氧化二砷对豚鼠心室肌细胞钾电流的影响.中国药学杂志,2008;43(24)1867-1870.
[89]高分飞,黄展勤,周燕琼,等.心肌细胞分离方法的改进及其钾电流的观察.广西医科大学学报,2008;25(3)338-340.
[90]陈修.心血管药理学[M].北京:人民卫生出版社,1989:483-486.
[91]Dorian P.Antiarrhythmic drug therapy of atrial fibrillation:focus on new agents.Cardiovasc Pharmacol,2003;8:S27-S31
[92]Peukert S,Brendel J,Pirard B,et al.Identification,synthesis and activity of novel blockers of the voltagegated potassium channel Kv1.5.J Med Chem,2003;46:486-498.
[93]Noda Y,Anzai K,Mori A,et al.Shirunei M and PackerL.Hyclroxyl and superoxide anion radical scavenging activitiesof natural source antioxidants using the computerized JES-FR30ESR spectrometer system.Biochem Mol Biol Int,1997;42(1):35-44.
[92]Pietri S,Maureli E,Drieu K,et al.Cardioprotective andantioxidant effects of the terpenoid constituents of ginkgo biloba extract(EGB761).J MolCell Cardio,1997;29(2):733-742.
[95]国家中医药管理局中华本草编委会.中华本草.第7期.第12卷,上海:上海科技出版社,1999,574.
[96]周玉新.中药指纹图谱研究技术,北京:化学工业出版社,2002:108.
[97]陈磊,康鲁平,秦路平,等.缬草属药用植物HPLC指纹特征研究.中药材,2003;26(4):256-259.
[98]石晋丽,刘勇,肖培根.缬草属药用植物极性成分的HPLC指纹图谱研究.中国中药杂志,2005;30(6):427-429.
[99]《中药注射剂指纹图谱研究的技术要求(暂行)的通知》.中国药品标准,2000;1(4):3.
[100]崔亚君,穆赫塔尔·亚森,岳松健,等.七种缬草属植物中缬草三酯和乙酰缬草三酯含量考察.西北药学杂志,1999;14(4):152-153.
[101]黄仁泉,张立,杨建丽.不同种缬草中缬草三酯和缬草烯酸类成分的HPLC的分析和比较.中草药,2002,33(11):1000-1001.
[102]傅亮,楚清脆,黄宝康,等.毛细管电泳一电化学检测法测定蜘蛛香中多元酚类化合物.分析化学,2005;33(2):161-164.
[103]明东升,郭济贤.缬草类生药中总缬草素的含量测定.上海医科大学学报,1993;20(3):210-212.
[104]陈磊,康鲁平,秦路平,等.总缬草素的质量标准和镇静催眠活性研究.中成药,2003;25(8):663-665.
[105]陈磊,秦路平,郑汉臣,等.三种缬草属植物的缬草素类含量种间和种内比较.中药材,2002;25(4):237-238.
[106]张素霞.缬草根和根茎中的挥发油、缬草酸及其衍生物和缬草三酯含量的季节变化及药用植株选择.国外医药.植物药分册,1999;14(1):21-22.
[107]左月明.黑水缬草抗老年痴呆有效部位化学成分和药理作用研究.黑龙江中医药大学博士学位论文,2007,5.
[108]中国药典.2005年版,一部,北京:人民卫生出版社,2005:附录33.