摘要
中药方剂是中医辨证论治、理法方药体系的重要组成部分,组方配伍是中药方剂研究的核心。随着医学模式和疾病谱的不断变化,需要更多新的中药方剂以应对越来越多的疾病挑战,中药方剂如何进行组方成为目前面临的关键问题,对中药方剂组方设计方法进行研究具有重要意义。
本文提出新中药方剂的组方设计方法并对其进行验证。在中医药理论指导下,选择中药经典名方,吸纳新的有效中药,化裁而出中药方剂基本方;将正交实验设计应用于中药方剂组方设计中的两个关键环节,即药味配伍和剂量配比。以典型的药理模型和特异性药效指标为判断标准,通过数学分析,在众多的比例中筛选出饮片层面的最佳药味配伍和剂量配比;运用化学分离和制剂分析技术,获得工艺稳定、质量可控的化学最佳制剂,再进行系统药效实验,验证其药效有效性,体现中药复方化学的整体研究理念;在此基础上进行系统药效学及药代动力学研究,探讨中药方剂作用、作用途径和中医药理论的相关性,推动方剂学科的发展。以治疗偏头痛的麻芎舒痛方为研究对象,对中药方剂组方设计方法进行验证,初步证明了中药方剂组方设计方法的可行性和科学性,为中药复方的配伍理论及应用提供有力支撑。
1基于药效学基础的麻芎舒痛方组方依据和剂量筛选
本文首先对中药复方的组方依据和剂量筛选进行药效学研究。以临床疗效为基础,以《中华人民共和国药典》为依据,拟定中药复方基本药味和用药范围。以镇痛效应为观察指标,通过正交试验,观察不同配比和不同剂量的中药复方药物对3种偏头痛动物模型的药效影响,筛选出中药复方药味配伍及最佳剂量配比。
1.1治疗偏头痛中药复方药物不同剂量配伍对硝酸甘油型偏头痛大鼠行为学的影响
观察不同药味配伍和不同剂量配比的中药复方对硝酸甘油型偏头痛大鼠挠头次数的影响。结果显示麻芎舒痛方(样本十)对偏头痛大鼠的挠头次数有最佳抑制作用。与模型对照组对比,统计学具有显著性差异(P<0.05)。
1.2治疗偏头痛中药复方药物不同剂量配伍对热板法小鼠镇痛作用的研究
观察在热板法小鼠模型实验中,不同药味配伍和不同剂量配比的中药复方的镇痛作用。结果显示(样本二、样本八和样本十)对小鼠热板法致痛时间有明显的延长效应。与模型对照组对比,统计学具有显著性差异(P<0.05)。
1.3治疗偏头痛中药复方药物不同剂量配伍对2%甲醛所致双相疼痛镇痛效应影响
观察不同药味配伍和不同剂量配比的中药复方的对2%甲醛所致双相疼痛镇痛效应的影响。结果显示样本六、样本七和样本十对2%甲醛致痛所产生的Ⅱ相疼痛时间有明显延长效应。与模型对照组对比,统计学具有显著性差异(P<0.05)。
综上所述,以3种动物模型的综合药理效应为实验依据,进行统计学和方差分析,筛选出包含治疗偏头痛的著名药对和经典药队的中药复方一麻芎舒痛方,为治疗偏头痛的中药复方提供了组方依据和优化剂量。
2不同治则配伍的中药方剂药效研究
采用硝酸甘油诱发的实验性偏头痛大鼠模型,以与偏头痛相关的5-HT作为药效学重要指标之一,观察不同治则组成的中药复方对大鼠脑组织和血清中的多种指标的影响,进一步确证麻芎舒痛方的药理作用。
对正常对照组、模型组、阳性药对照组和中药复方治则配伍组在偏头痛大鼠血清中5-HT、CGRP和ET-1的含量进行统计分析,结果表明:与模型组相比,天麻、川芎配伍组可以显著降低偏头痛大鼠血清中5-HT含量(P<0.05)。麻芎舒通方水提物可以显著降低偏头痛大鼠血清中5-HT和ET-1含量(P<0.05)。麻芎舒通方醇提物可以显著降低偏头痛大鼠血清中CGRP、ET-1和5-HT含量(P<0.05)。
对正常对照组、模型组、阳性药对照组和中药复方治则配伍组在偏头痛大鼠脑组织中5-HT含量进行统计分析,结果表明:与模型对照组相比,天麻川芎配伍组、麻芎舒通方水提物和麻芎舒通方醇提物均可以显著降低偏头痛大鼠脑组织中在5-HT含量(P<0.05)。
对不同治则配伍的中药方剂进行药效研究,结果显示:以平肝降火、活血行气、散通络止痛为治则的麻芎舒通方药效优于天麻、川芎配伍组。不同的提取工艺具有不同的药理效应,麻芎舒通方乙醇提取优于水煎提取。
3中药复方(麻芎舒痛方)的药学研究
如何正确地评价中药制备工艺的合理与优劣,是目前困扰中药研究的问题之一。中药复方药学研究是中药复方研究的基础,尽可能地将发挥药效和发挥疗效的物质基础提取出来,才能更好地运用于临床,服务于患者。最佳制备工艺研究是中药复方药学研究的关键环节。
3.1川芎不同提取部位治疗偏头痛的药效学研究
本文在比较关键的药物或不好确定药物提取路线时,以药效学试验为依据,对川芎多种提取物进行比较,将中药化学-药理效应相结合,并采用HPLC-ECD技术测定偏头痛大鼠血液和脑组织中的5-HT含量,最终确定川芎乙醇提取;通过对其余四味药物化学成分和药理成果的分析,确定四味药物醇提。
3.2中药复方提取路线和提取方式的确定
以君臣药物的有效成分为指标,对复方的混合提取方式和单独提取方式进行考察,天麻素混和提取/单独提取的比值是102.42%,葛根素混和提取/单独提取的比值是108.54%,说明混合提取更有利于天麻和葛根中有效成分的提取。阿魏酸混和提取/单独提取的比值是99.46%,说明对于阿魏酸来说,混合提取和单独提取基本相当。从大生产和成本考虑,川芎亦选择混合提取方式。
3.3麻芎舒痛方的制备工艺研究
以L_9(3~4)正交试验进行最佳工艺条件的筛选和验证,确定了复方的最佳制备工艺,即五味药物以8倍量60%的乙醇提取3次,每次2小时。此工艺在保证君药有效成分提取的同时,兼顾了两味臣药有效成分的提取。五味药物同时提取,具有成本降低、工艺简单有效之优点,有利于大生产。
4中药复方(麻芎舒痛方)的制剂质量控制
4.1仪器与试药
高效液相色谱仪:Waters600泵,Waters 996检测器,Waters717自动进样器,Millennium~(32) Login工作站;薄层扫描仪,型号:TRANSILLUMINATOR 2020D;空气压缩泵,TOSHIBA Corporation生产;阿魏酸、天麻素、葛根素、川芎对照药材和白芷对照药材为对照品或对照药材。
4.2 TLC色谱鉴别
照中国药典薄层色谱法(附录ⅥB)试验,对川芎、天麻和葛根进行TLC定性鉴别。川芎的薄层色谱鉴别采用川芎对照药材,以正己烷.醋酸乙酯(9∶1)为展开剂,供试品色谱中,在与对照药材色谱相应的位置上,显相同颜色的荧光斑点;葛根的薄层色谱鉴别采用葛根素对照品,以氯仿-甲醇-水(7∶2.5∶0.25)为展开剂,供试品色谱中,在与对照品色谱相应的位置上,显相同颜色的荧光斑点;白芷的薄层鉴别采用白芷对照药材,以石油醚(60-90℃)-乙醚(1∶1)为展开剂,。供试品色谱中,在与对照品色谱相应的位置上,显相同颜色的荧光斑点。阴性对照均无干扰。
4.3含量测定
4.3.1样品中阿魏酸的含量测定
色谱柱:SymmetryC18(3.9×150 mm,5μm);流动相A:乙腈-0.05%醋酸水溶液,流动相B:甲醇。0～45 min,A由100%至60%,B由0%至40%,45～50 min,A由60%至0%,B由40%至100%;检测波长为323 nm;流速:1.0ml·min~(-1);柱温:30℃;阿魏酸在0.02715～0.4887μg范围内,峰面积和进样量呈良好的线性关系(γ=0.9998,n=7);阿魏酸样品回收率为98.22%,RSD为1.30%(n=6);本品每1g含川芎以阿魏酸(C_(10)H_(10)O_4)计,不得少于0.18 mg。
4.3.2样品中天麻素的含量测定
色谱柱:Symmetry C18(3.9×150 mm,5μm);流动相:乙腈-0.05%冰醋酸水溶液(2:98);检测波长为220nm;流速为0.6 ml·min~(-1);柱温:室温;天麻素在0.1032～1.0320μg范围内,峰面积和进样量呈良好的线性关系(γ=1.0000,n=7);天麻素样品回收率为97.81%,RSD为1.49%(n=6);本品每1g含天麻以天麻素(C_(13)H_(18)O_7)计,不得少于1.92 mg。
4.3.3样品中葛根素的含量测定
色谱柱:SymmetryC18(3.9×150 mm,5μm);流动相A:乙腈-0.05%醋酸水溶液,流动相B:甲醇。0～45min,A由100%～60%,B由0%～40%,45～69min,A由60%～0%,B由40%～100%;检测波长为250nm;流速:1.0 ml·min~(-1);柱温:30℃;葛根素在0.164～2.05μg范围内,峰面积和进样量呈良好的线性关系(γ=0.9998,n=9);葛根素样品回收率为97.87%,RSD为1.57%(n=6);本品每1g含葛根以葛根素(C_(21)H_(20)O_9)计,不得少于13.18 mg。
5中药复方中多成分在体肠吸收研究
本文初步考察了中药复方中川芎、天麻、葛根的有效成分阿魏酸、天麻素和葛根素在体肠吸收行为。复方配伍和单味药材的肠吸收行为进行比较,阿魏酸的单位面积肠吸收量增加,天麻素和葛根素的单位面积肠吸收量减少,体现出中药复方配伍的协同或拮抗作用;当药物浓度增大时,阿魏酸的单位面积肠吸收量基本相当,天麻素和葛根素的单位面积肠吸收量随浓度增大而增加,药物的吸收属于被动吸收范畴;基于天麻素在复方配伍时天麻素的肠吸收量有较大降低,故将不同药物与天麻进行配伍研究,旨在寻求抑制天麻素吸收的药材来源。天麻分别与川芎、白芷配伍时,天麻素的肠吸收率均达到90%左右。而天麻与葛根配伍时,天麻素的吸收率为46%,天麻素的肠吸收有较大程度的降低,说明复方配伍时抑制天麻素肠吸收的药物是葛根。
6 LC-MS-MS法对中药复方中多种有效成分的定性研究
本文应用LC-MS-MS法对中药复方(麻芎舒痛方)中多种有效成分进行定性研究。建立了多种成分于同一液相色谱图中的梯度洗脱系统,以LC-MS-MS负离子扫描方式对复方样品Krebes-Ringer's液中的多种成分进行定性研究,共明确了川芎和葛根中的8种成分,分别为川芎中的阿魏酸、葛根中的4′,7-二葡萄糖苷大豆苷元、3′-羟基葛根素、葛根素、3′-甲氧基葛根素、葛根素木糖苷、大豆苷、大豆黄素。
在定性研究中推测出结构的一种成分是滨蒿内酯(Scoparone),推测它有可能是麻芎舒痛方在共同的提取过程中产生的一种新成分。查阅文献滨蒿内酯的植物来源多为菊科植物滨蒿Artemisia scoparia Waldst.et Kit.花、种子,茵陈蒿A.capillaris Thunb.茎、叶,芸香科植物印度枸(木桔)Aegle marmelos Corr.根和伞形科植物木柴胡Bupleurumfruticescens L.地上部,本复方中没有这些植物,滨蒿内酯(Scoparone)的来源需要进一步确认。LC-MS-MS法对中药复方的定性研究为其物质基础研究和更深入的体内代谢研究打下基础。
7研究的创新性
◆提出“将多种药理模型与特异性指标相结合,以药味配伍和剂量配比为筛选依据”的中药方剂组方设计研究方法,并对麻芎舒痛方的药味配伍和剂量配比进行筛选,验证了组方设计的可行性和合理性;
◆选择与疾病相关的特异性药效指标成分,以最佳制剂进行药效试验验证和药代动力学探索性研究,探讨中药方剂作用和中药复方配伍的相关性;
◆建立了多种成分于同一液相色谱图中的梯度洗脱系统,探讨麻芎舒痛方中多成分的在体肠吸收行为,对麻芎舒痛方药物相互配伍规律进行探索性研究,为进一步的药代研究打下基础。
Design Studies of the Fractional Optimization of TCM Formula(MXST)
A Traditional Chinese Medical formula(TCM formula) is one of the most important parts of the Chinese medical science.Studies on composing prescriptions and drug compatibility are the core of TCM formulas.With the changes of medical mode,more and more new TCM formulas are required.It is the key question to design the prescriptions.
This paper puts forward and verifies a new design method of new TCM formulas.Orthogonal experiment design is used during the formulation design of a new TCM formula which is composed of a classical prescription and effective TCM.Two key links in a new TCM formula are the best dose design and drug compatibility,which is obtained by using classical animal model and special pharmacology index; The best chemical preparation is obtained by using the chemical and analysis techniques.System studies on the pharmacology embody the whole view of chemistry of compound prescriptions.System researches on the pharmacology and Pharmacokinetics and the correlation with TCM theory promote the subject development of the TCM prescription.By taking MXSTF as a research object,exploratory researches on TCM formula prove the feasibility and scientific.
1 The base of prescription formulation and dose screening on the basis of pharmacodynamics of MXSTF
Formula screening by the research of pharmacodynamics was studied firstly.The basic recipe is drafted based on the clinical efficacy and PRC codex.Analgesic effect was the observation index by the orthogonal design.Efficacy impact was studied by TCM formulas with different proportions and doses on three animal models of migraine,then the best drug compatibility and dose ratio were screened.
1.1 Influences on the model rats of migraine induced by nitroglycerin by TCM formulas with different proportions and doses
The influences on the model rats of migraine induced by nitroglycerin by TCM formulas with different proportions and doses were observed.Results:The tenth sample(MXSTF) has the best analgesic effect on the model rats of migraine induced by nitroglycerin.Compared to the model control group,it is showed statistical difference(P<0.05).
1.2 Studies on the pain time of model mouse induced by hot plate by TCM formulas with different proportions and doses
The studies on the pain time of model mouse induced by hot plate by TCM formulas with different proportions and doses were observed.Results:Pain Time was obviously postponed on the model mouse induced by hot plate because of the second sample、the eighth sample and the tenth sample.Compared to the model control group,it is showed statistical difference(P<0.05).
1.3 Influences on the delay time of model mouse of induced by 2%formaldehyde by TCM formulas with different proportions and doses
The influences on the delay time of model mouse of induced by 2%formaldehyde by TCM formulas with different proportions and doses were observed.Results:Delay time was obviously longer then other samples on the model mouse ofⅡ-phase-pain induced by 2%formaldehyde because of the sixth sample、 the seventh sample and the tenth sample.It is showed statistical difference when compared to the model control group(P<0.05).
In a word,the base of prescription formulation and dose screening of the TCM formula curing migraine were set up according to these pharmacological effects,which include the famous drug pair and classical drug team.
2 Studies on the pharmacology of TCM formulas with different therapeutic principle compatibility
The influence on the contents of index components was observed by TCM formulas with different therapeutic principle compatibility in rat's brain tissue and serum.The model rats of migraine induced by nitroglycerin and special pharmacology index were used during the experiment in order to verify the pharmacological action of MXSTF.
The contents of 5-HT、CGRP and ET-1 in rat's serum were statistically analyzed among normal control group、positive drug control group and TCM formulas groups with different therapeutic principle compatibility.Compared with model group,there were significant differences in 5-HT level from the group of Rhizoma Gastrodiae and Rhizoma Chuanxiong(P<0.05),in 5-HT and ET-1 levels from MXSTF water extract(P<0.05),and in 5-HT、ET-1和CGRP levels from MXSTF alcohol extraction(P<0.05).
The levels of 5-HT in rat's brain tissue were statistically analyzed among normal control group、positive drug control group and TCM formulas groups with different therapeutic principle compatibility. The results show that there were obvious differences among Rhizoma Gastrodiae and Rhizoma Chuanxiong group、MXSTF water extract and MXSTF alcohol extraction when compared with model group(P<0.05 ).
The pharmacological actions show that MXSTF is superior to Rhizoma Gastrodiae and Rhizoma Chuanxiong group and MXSTF alcohol extraction is better than MXSTF water extract.
3 Studies on the pharmacy of MXSTF
The pharmacy studies is the base of the study of TCM formula.In order to improve clinical effect,the main material basis of drug must be extracted as much as possible.Technology is a key link to the study on the pharmacy of TCM formula.It is one of the issues that how to correctly evaluate the preparation process of Chinese medicine.
3.1 Effects on the pharmacodynamic of different ingredients extracted from Chuanxiong
Pharmacodynamic experiments were as indexes when the route of drug extraction is difficult to determine.Many ingredients were extracted from Chuanxiong.All of them were compared with the pharmacodynamics of migraine.The mode with the combination of Chinese Medicine Chemistry and Pharmacology of TCM is used to determine the extraction route of Chuanxiong.The content of 5-HT was determined by electrochemical method during migraine.Then chuanxiong is extracted by ethanol;Other drugs were also extracted by ethanol.
3.2 Determination of the extraction route and the extraction way
Using effective components of monarch drug and ministerial drugs as a parameter,the effects of mixed extracting and separate extracting were observed.The ratio of mixed extracting/separate extracting for Gastrodin is 102.42%,the ratio of mixed extracting/separate extracting for Puerarin is 108.54%.The dates showed that mixed extracting is more beneficial to the extraction of Tianma and Gegen.The ratio of mixed extracting/separate extracting for Ferulic acid is 99.46%.The mixed extracting is equal to separate extracting for Ferulic acid,so the mixed extracting was chosen for Chuanxiong.
3.3 Studies on the technology of MXSTF
The optimum mixture extraction process by L_9(3~4) is as follows:80%alcohol extracting third,adding 8times amount of alcohol into Chinese medical materials,once 2 hours.The method has features of low cost,simple process and effectiveness,which will be advantageous to extend and use the formula.
4 Quality control of TCM formula(MXSTF)
4.1 Instrument and testing drug
HPLC:Waters 600 pump,Waters 996 detertor,Waters 717 auto sampler,Millennium~(32) Login;TLC scanner,TRANSILLUMINATOR 2020D;air compressor pump,TOSHIBA Corporation.
Reference substance:Ferulic acid,Gastrodin,Puerarin;Reference drugs:Rhizoma Chuanxiong,Radix Angelicae Dahuricae.
4.2 TLC chromatographic identification
n-Hexane-ethyl acetate(9:1) was used for Rhizoma Chuanxiong with crude standard as reference; Trichloromethane- methanol-water(7:2.5:0.25) was used for Radix puerariae lobatae;Petroleum ether-Ether(1:1) was used for Radix Angelicae Dahuricae with crude standard as reference.
4.3 Determination
4.3.1 Determination of Ferulic acid in sample
Column:SymmetryC_(18)(3.9×150 mm,5μm);The mobile phase A was acetonitrile-0.05%acetic water, the mobile phase B was methanol.0-45 min,A from 100%to 60%,B from 0%to 40%;45 - 50 min,A from 60%to 0%,B from 40%to 100%;The detection wavelength was 323 nm;Column temperature was 30℃;The flow rate was 1.0 ml·min~(-1);A good linearity of Ferulic acid presented over the range of 0.02715-0.4887μg(γ=0.9998,n=7);The average recovery rate for the content determination was up to 98.22%,RSD=1.30%(n=5);Determination of Ferulic acid in the preparation was not less than 0.18 mg per one gram.
4.3.2 Determination of Gastrodin in sample
Column:SymmetryC_(18)(3.9×150 mm,5μm);The mobile phase was acetonitrile-0.05%acetic water (2:98);The detection wavelength was 220 nm;The flow rate was 0.6 ml·min~(-1);Column temperature was room temperature;A good linearity of Gastrodin presented over the range of 0.1032-1.0320μg(γ=1.0000, n=7);The average recovery rate for the content determination was up to 97.81%,RSD=1.49%(n=6); Determination of Gastrodin in the preparation was not less than 1.92 mg per one gram.
4.3.3 Determination of Puerarin in sample
Column:SymmetryC_(18)(3.9×150mm,5μm);The mobile phase A was acetonitrile-0.05%acetic water, the mobile phase B was methanol.0-45 min,A from 100%to 60%,B from 0%to 40%;45 - 69 min,A from 60%to 0%,B from 40%to 100%;The detection wavelength was 250 nm;Column temperature was 30℃;The flow rate was 1.0 ml·min~(-1);A good linearity of Puerarin presented over the range of 0.164-2.05μg(γ=0.9998,n=9);The average recovery rate for the content determination was up to 97.87%, RSD=1.57%(n=6);Determination of Puerarin in the preparation was not less than 13.18 mg per one gram.
5 Studies on Absorption Kinetics of Multi-composition by Intestinal Backflow in vivo
Intestinal absorption behaviors of Ferulic acid、Gastrodin and Puerarin were observed in this thesis. Compared to single herb,the uptakes amount per unit area of Ferulic acid increase in the TCM formula, while the uptake amounts of Gastrodin and Puerarin per unit area decrease.Synergistic or antagonistic effect was shown between compatibility in TCM formula.With drug concentration increasing,the uptake amounts of Gastrodin and Puerarin were increased.The absorption way fits the passive transport mechanism;Based on the great decrease of Gastrodin in TCM formula,the studies of compatibility of Tianma with different drugs were carried out.The absorption rate of Chuanxiong and Tianma、Baizhi and Tianma were all for 90%,while the absorption rate of Gengen and Tianma was 46%.Therefore Gengen is the herb source which restrains the absorption of Gastrodin in intestinal.
6 Qualitative Study on Multi-composition in TCM Formula by LC-MS-MS
This paper established gradient elution system which can include multi-composition in the same chromatogram.Eight kinds of elements of Chuanxiong and Gengen dissolved in Krebes -Ringer's solution were determined with LC-MS-MS negative ion scan mode,which are Ferulic acid、daidzein 4'-7-diglucoside、3'-OH Puerarin、Puerarin、3'-OCH_3 Puerarin、Puerarin xyloside、Daidzin and Daidzein respectively.
It needs to further research on two unknown materials and Scoparone.The source of Scoparone needs to be further determined because the TCM formula do not include the plants which Scoparone coming from,namely Artemisia scoparia Waldst.et Kit.、A.capillaris Thunb.、Aegle marmelos Corr.and Bupleurum fruticescens L.Qualitative study by LC-MS-MS is as a basis for further studies on the material base and metabolism in vivo.
引文
[1]于彪.川芎对药治疗头痛的临床应用.中医药学报,2001,29(2):11.
[2]彭怀仁.中医方剂大辞典.北京:人民卫生出版社,1993.
[3]王洪喜,丁元庆.天麻治疗头痛的药对配伍规律.山东中医杂志,2006,25(2):101-103.
[4]段重高,李庶伟.葛根素对金黄地鼠脑微循环的影响.中华医学杂志,1991,71(9):516.
[5]卢冬初.葛根对蛛网膜下腔出血的剧烈头痛之疗效.江西中医药,1996增刊,19.
[6]Xu Xiaohong,Zhang Shaomin,Zhang Lei,et al.The Neuro-protection of puerarin against cerebral ischemia is associated with the prevention ofapoptosis in rats.Plantamedica,2005,71(7):585-591
[7]唐文炳.葛根素尼莫地平治疗血管性头痛对比研究.医药论坛杂志,2006,27(23)75-77.
[8]孙明谨,高才兵,李雪锋,等.葛根素及尼莫地平治疗血管性头痛疗效对比研究.实用诊断与治疗杂志,2007,21(10):745-747.
[9]陈珺.葛根汤联合行为干预治疗紧张性头痛临床观察.中国中医急症,2008,17(2):160-161.
[10]王林森.延胡索在痛证中的应用.中国中医基础医学杂志,1998,4(8):197-198.
[11]黄欣,张敏,苏乐群,等.元胡止痛制剂研究概述.山东中医杂志,2005,24(1):43-45.
[12]陈国华,孙阔,张社卿.元胡止痛片治疗偏头痛176例.光明中医,2003,18(5):46-47.
[13]Saxena PRXranial arteriovenous shunting,an in vivo animal model for migraine.Experimental Headache Models,1995:189-198.
[14]Tassorelli C,Nappi G,Joseph SA.Systemic nitroglycerin induces Fos immunoreactivity in rat brain.Experimental Headache Models,1995:229-234.
[15]Moskowitz MA.Neurogenic versus vascular mechanisms of suma-triptan and ergot alkaloids in migraine.Trends PharmacolSci,1992,13(8):307-31.
[16]陈磊,王怀良,邢军,等.植物药生物碱94-95-10L对大鼠CSD模型脑内c-fos基因表达的影响.中国医科大学学报,2000,29(3):172-174.
[17]杜力平.精制吴茱萸胶囊对偏头痛小鼠作用的研究.中药药理与临床,1998,14(5):4-6.
[1]李小玲,李德炎.五羟色胺影响的行为谱.国外医学(精神学分册),1999,26(3):172-175.
[2]钟华,王玲玲,韩振蕴,等.脑痛立停胶囊对偏头痛模型大鼠血浆CGRP及ET的影响,北京中医药,2008,27(8):656-658.
[3]Ottosson A,Edvinsson L.Release of histamine from duralmast cells by substance Pand cal-citoningene-related peptide[J].Cephalalgia,1997,17(3):166-172.
[4]彭良富,杨期明,廖远高.偏头痛患者血浆内皮素-1含量的变化研究.华夏医学,2006,19(2):252-253.
[5]田青,孙国杰.针刺对偏头痛患者血浆内皮素水平的调节.上海针灸杂志,1998,17(2):5.
[1]中华人民共和国药典委员会.中国药典2005年版一部.化学工业出版社,2005年
[2]北京制药工业研究所.川芎的化学成分研究.北京医药工业,1975,1-5.
[3]袁伯勇,袁慧,赵铭.3种饮片中川芎嗪含量的测定.中草药,2000:31(6):430-431.
[4]杨广德,梁明金,贺浪冲,等.川芎中阿魏酸的提取方法研究.中成药,2002,24(6):418-420.
[5]黄雪梅,罗淑萍.RP-HPLC法测定龙胆泻肝口服液中阿魏酸的含量.中国实验方剂学杂志,2001,7(3):3-4.
[6]江滨,许晓峰,韦群辉.滇藁本精油中镇静成分的分离与鉴定.中国民族民间医药杂志,1998,33:3.
[7]石力夫,郑晓梅,秦溱,等.藁本内酯分解前后川芎挥发油对兔球结膜微循环影响的比较.中国药理学与毒理学杂志,1995,9(2):157.
[8]王浴生.中药药理与应用.北京:人民卫生出版社,1983.P119.
[9]张华,雒晓东,连新福,等.天麻素临床应用研究进展.光明中医,2006,21(12):70-72.
[10]任东波.天麻连续给药对小鼠的镇痛作用.农业与技术,1999,19(5):74.
[11]董英,徐斌,林琳,等.葛根的化学成分研究.食品与机械,2005,21(6):85-88.
[12]岳红文,胡小琴.葛根及葛根素对心血管系统的药用价值.中国中西医结合杂志,1996,16(6):382.
[13]杨金云.葛根抗心率失常研究.中国现代药理杂志,1998,(2):53.
[14]王晓青,傅静.葛根的药理作用研究进展.北京中医药大学学报,1994,17(3):39.
[15]王梦月,贾敏如,马逾英,等.白芷镇痛有效部位的化学成分研究.中国药学杂志,2005,40(8):583-585.
[16]吕清文,丛雅勤.延胡索药理作用及临床应用浅议.内蒙古中医药,2004,9:26.
[17]林青,付文卫,李江,等.茱萸汤的复方化学研究.贵阳中医学院学报,2000,22(1):57-59.
[18]袁久荣,李以凤,袁浩.四物汤的实验研究.中国中药杂志,1991,16(3):153-155
[19]朱丹妮,严永清,李志明.生脉散复方化学动态变化与药效关系的研究-生脉散复方化学的研究(Ⅰ).中国中药杂志,1998,23(4):230-232.
[20]朱丹妮,李志明,严永清,等.生脉散复方化学动态变化与药效关系的研究-生脉散复方化学的研究(Ⅱ).中国中药杂志,1998,23(5):291-293.
[21]李秀玲,肖红斌,徐青,等.中药川芎和赤芍配伍规律的方法研究.高等学校化学学报,2004,25(9):1632-1636.
[22]陈静,刘世喜,胡国华,等.天麻素注射液治疗眩晕的多中心随机单盲对照试验.中国循证医学杂志,2004,4(12):864-870.
[1]程锦,狄留庆.在体肠段灌流模型在中药吸收研究中的应用.Chinese Journal of Information on TCM,2008,15(2):98-100.
[2]黄沛,杨中林.不同纯度葛根素提取物的大鼠小肠吸收动力学研究.海峡药学,2008,20(3):43-45.
[3]洪震,曾军,黄茂盛,等.缺血性和出血性脑卒中的发病率与年龄的关系.上海医学,2000,23:663.
[4]郭歆,曹伟,程泽能,等.齐墩果酸大鼠肠吸收动力学.中南药学,2007,5(3):216-219.
[5]Thrift AG,Dewey HN,Macdonell RA,et al.Stroke incidence on the east coast of Australia:the north east Melbourne stroke incidence study(NEMESIS).Stroke,2000,31:2087.
[6]谭晓斌,贾晓斌.刺五加苷B的大鼠在体肠吸收特性研究.中成药,2008,30(3):346-349.
[7]聂淑芳,潘卫三,杨星刚,等.对大鼠在体单向灌流技术中重量法的评价.中国新药杂志,2005,14(10):1176-1179.
[1]国家医药管理局中草药情报中心站.植物药有效成分手册.北京:人民卫生出版社
[2]董英,徐斌,林琳,等.葛根的化学成分研究.食品与机械,2005,21(6):85-88.
[1]姜廷良.试论中药方剂的研究与发展.中药药理与临床研究进展,1996,4:16.
[2]林青,付文卫,李江,等.茱萸汤的复方化学研究.贵阳中医学院学报,2000,22(1):57-59.
[3]袁久荣,李以凤,袁浩.四物汤的实验研究.中国中药杂志,1991,16(3):153-155.
[4]朱永新,严克东,涂国士.复方制剂生脉散的化学分析研究.药物分析杂志,1989,9(1):5-9.
[5]严永清,朱丹妮,吴建新,等.六味地黄汤化学研究方法的探索.中国中药杂志,1991,16(5):310-312.
[6]罗尚凤,李国光,何宇东,等.枳术丸的化学成分分析.西北药学杂志,1994,9(5):206-209.
[7]罗尚凤,朱俊峰,武春华,等.枳术丸组方与炮制工艺的研究.中国中药杂志,1994,19(1):23-24.
[8]朱丹妮,严永清,李志明.生脉散复方化学动态变化与药效关系的研究-生脉散复方化学的研究(Ⅰ).中国中药杂志,1998,23(4):230-232.
[9]朱丹妮,李志明,严永清,等.生脉散复方化学动态变化与药效关系的研究-生脉散复方化学的研究(Ⅱ).中国中药杂志,1998,23(5):291-293.
[10]李秀玲,肖红斌,徐青,等.中药川芎和赤芍配伍规律的方法研究.高等学校化学学报,2004,25(9):1632-1636.
[11]裴妙荣,梁秀和,廖辉,等.薄层扫描法对四逆汤配伍作用的化学分析.中成药,1993,15(6):33-35.
[12]陈建萍,谭炳炎,张敏生,等.四逆汤中附子甘草配伍规律研究.中国实验方剂学杂志,2001,7(3):16-17.
[13]裴妙荣,王世民,李晶.四逆汤中甘草对附子解毒作用的相关性分析.中国中药杂志,1996,21(1):50-53.
[14]张爱华,彭国平,文红梅,等.甘草与附子配伍煎液的甘草黄酮含量测定.中成药,1999,21(4):196-198.
[15]何燕萍.复方中成药慢肝宝冲剂中五昧子乙素的含量测定.中国中药杂志,1995,20(9):541-543.
[16]方洪,苏子仁,梁永枢,等.复方虎茵汤方药配伍对虎杖有效成分煎出的影响.中国实验方剂学杂志,1997,3(5):1-2.
[17]何桂霞,蒋孟良,冯映冰.芍药汤配伍的化学变化研究.中国中药杂志1998,23(7):432-434.
[18]HuangH,LiangMJ,ZhangXM,etal.Simultaneous determina-tion ofnine flavonoids and qualitative evaluation ofHerbaEpimedii by high performance liquid chromatographywith ultravioletde-tection.J Sep Sci,2007,30:3207.
[19]尹莲,徐立,时东.加味四妙丸有效部位群的筛选研究思路与方法.2005,7(4):28.
[1]郭东艳,杨大坚,陈士林.葛根素及其衍生物在大鼠体内的药代动力学研究.陕西中医学院学报,2008,31(1):52-54.
[2]田景振,魏凤环.葛根黄酮缓释胶囊家兔体内药代动力学研究.山东中医药大学学报,2003,27(6):454.
[3]杜力军,於兰,常琪,等.新工艺制备的葛根黄酮的药代动力学研究.中药药理与临床1997,13(2):19-24.
[4]谭晓梅,吴艳萍.葛根芩连汤配伍葛根素在兔体内药动学的研究.中药药理与临床2006,22(5):1-3.
[5]卢弘,邢东明,孙虹,等.CBN中葛根素、人参皂甙Rgl在正常和脑缺血再灌注大鼠体内药代动力学比较.哈尔滨商业大学学报(自然科学版),2001,3:5.
[6]王雪莉,邢东明,盖国胜,等.微细化工艺制备的葛根微粉溶出度和生物利用度的研究.中国药学杂志,2004,39(4):283.
[7]张志荣,游学均,何勤,等.HPLC测定血浆中葛根素.中国药学杂志,1997,32(2):104-106.
[8]李妮,路玫,陈晖,等.葛根素在糖尿病肾病病人中的药动学.中国新药与临床杂志,2002,21(1):51.
[9]郭炎荣,蒙大平,韦戈,等.高效液相色谱法测定全血中葛根素含量.中国医院药学杂志,2000,20(6):343.
[10]邓新国,胡世兴,张清炯,等.葛根素腹腔注射在家兔房水中的药代动力学研究.眼科研究,2003,21(3):258.
[11]王莉,邢尔明,卢弘,等.CBN中有效成分葛根素在大鼠体内的排泄.中药药理与临床,2001,17(5):8-10
[12]郭莹,万海同,张莉.不同中约复方有效部位组合对脑缺血大鼠血浆中葛根素代谢变化的研究.中国中药杂志,2007,32(23):2531-2534.
[13]郭建平,李国栋,赵彤英,等.HPLC对人血浆中葛根素及葛根黄酮的线性考察.药学实践杂志,2003,21(4):219.
[14]张莉,黄熙,任平等.三维HPLC法同步测定犬血浆中的葛根素及阿魏酸.沈阳药科大学学报,2001,18(1):45.
[15]黄熙,任平,张莉,等.HPLC直接测定血清阿魏酸-方剂血样预处理新方法.中草药,1999,30(3):175.
[16]马娜,陈玲,熊飞.固相萃取技术及其研究进展.上海环境科学,2002,21(3):181-184.
[17]王莉,邢东明,孙虹,等.金森脑泰注射剂中葛根素在生理及病理状态下大鼠肝肾分布的比较.中国实验方剂学杂志,2002,8(3):41.
[18]黄熙,任平,张莉,等.HPLC法测定犬口服养阴通脑颗粒后血浆中的葛根素.中国药科大学学报,2001,32(2):127-129.
[19]金昔陆,朱秀媛,王文杰等.血浆中葛根素的高效液相色谱测定法及其在狗体内的药代动力学.药学学报,1997,32(10):782.
[20]朱秀媛,苏成业,李振华,等.葛根有效成分的代谢研究Ⅲ.葛根素的代谢及其药代动力学分析.药学学报,1979,14(6):349.
[21]金昔陆,朱秀媛.血浆中葛根素的薄层-紫外光密度扫描测定法及其在家兔的药动学.中国药理学通报,1991,7:421.
[22]王健,赵岚峰,王杰,等.高效毛细管电泳简介及其在中药分析中的应用.中成药,1996,18(10):45.
[23]杜力军,邢东明,炎彬,等.对葛根素与葛根黄酮体内动力学关系的探讨.世界科学技术a) -中医药现代化(基础研究),2004,6(6):26-30.
[24]刘昌孝.药物代谢动力学.长沙:湖南科学出版社,1979.P174.
[25]王成,刘玉玲,谷士杰.葛根素的溶解性及络合助溶的研究.中国药学杂志,1993,28(5): 294.
[26]刘振威,陈雪梅,高尔.磷脂对大鼠离体肠管吸收葛根素的影响.潍坊医学院学报,1998,20(4):253.
[27]刘振威,潘爱美,李公宝,等.磷脂对葛根素改善家兔血液流变学和微循环作用的影响.潍坊医学院学报,1998,20(4):250.
[28]林文慧,朱春燕,陈卫.葛根黄酮在大鼠肠道的吸收动力学研究.中国中药杂志,2008,33(2):164-168.
[29]周冬菊,赵会英,杨英禄.大鼠小肠对葛根素吸收的动力学研究.北京化工大学学报,2006,33(5):106.
[30]郭建平,成国祥,孙其荣,等.葛根黄酮滴丸兔体内药动学.中国药学杂志,2004,39(5):364-365.
[31]金昔陆,程桂芳,朱秀媛.葛根素在健康志愿者的药代动力学.中国临床药理学杂志,1991,7(2):115-118.
[32]吴正红,朱延勤,程光忻,等.葛根素滴眼液眼内药代动力学的研究.中国药科大学学报,1998,29(3):221.
[33]严汉英,朱延勤,吴正红,等.葛根素滴眼液降眼压作用及其在眼房水中的药物浓度.眼科新进展,1999,19(2):73.
[34]金昔陆,朱秀媛.葛根素在三种动物的药代动力学参数与体重的相关性.徐州医学院学报,1991,11(1):25.
[35]崔升淼,赵春顺.大鼠离体肝灌流中葛根素的代谢动力学研究.中药药理与临床,2008,24(1):17-19.
[36]黄华,王显著.表面活性剂对葛根素固体分散体体外溶出的影响.中国药科大学学报,2004,35(4):315-317.
[37]李彩君,王燕玲,黄颖然.薄荷醇对葛根素体外渗透动力学的影响.药物研究,2007,4(18):184-185.
[38]刘晓彬,权丽辉,刘春雨,等.甲基化壳聚糖对葛根素小鼠口服吸收的影响.世界科学技术—中医药现代化(基础研究),2007,9(6):51-54.
[39]王锦,李颖,杨丽彬.氮酮对葛根素透皮吸收作用的实验研究.药学实践杂志,1995,13(2):93-95.
[40]李彩君,王燕玲,方馥蕊.辛类中药挥发油对葛根素的透皮吸收促进作用研究.药物研究,2007,4(22):141-142.
[41]朱春燕,王雪莉,石峰.葛根总黄酮缓释片药动学研究.世界科学技术—中医药现代化(基础研究),2007,9(6):58-60.