【作者】 王光忠；

【导师】 刘焱文；

【作者基本信息】 湖北中医学院 ， 中医基础理论， 2009， 博士

【摘要】 脑得生片（丸）为传统中药制剂，收载于《中国药典》2005年版一部，由三七、川芎、红花、葛根、山楂五味中药组成，具有活血化瘀，通经活络之功效。用于瘀血阻络所致的眩晕、中风，症见肢体不用、言语不利及头晕目眩；脑动脉硬化、缺血性中风及脑出血后遗症。临床应用广泛，疗效确切。本研究论文在抗血栓药理活性指导下，对复方脑得生药效物质的提取工艺、化学成分、含量分析、指纹图谱、入血成分等进行了相关研究，旨在为脑得生制剂的二次开发提供科学依据。采用超临界CO₂萃取技术对川芎有效部位的提取工艺进行了研究，获得了可靠的技术参数，进行了中试验证，可过渡到产业化生产。采用中药药理学方法，对川芎药材不同部位进行了抗血栓活性的药理筛选研究，结果表明，川芎超临界CO₂萃取物对大鼠血栓湿重显著减轻，其作用优于川芎总提物，说明采用超临界CO₂萃取技术应用于川芎提取物的生产工艺具有可行性。采用GC-MS法，对其有效部位（川芎油）的药效物质成分进行了分析，分离并鉴定了35种成分。采用GC法建立了川芎油的指纹图谱，共标定了26个共有峰，并结合GC-MS法对各共有峰进行了鉴定，方法的重现性和稳定性良好，从而为川芎药材及其提取物建立与国际接轨的质量评价标准奠定了基础。采用溶剂法和大孔树脂纯化技术，对复方脑得生主要有效部位总黄酮的提取纯化工艺进行了优选研究，获得了可靠的技术参数，进行了中试验证，可过渡到产业化。采用分光光度法和高效液相色谱法，分别对提取物中的总黄酮及其主要活性成分葛根素和羟基红花黄色素A进行了含量分析研究，建立了重现性好、可靠性强的含量测定方法，能有效控制该复方总黄酮提取物的质量。采用高效液相色谱法，对提取物进行了指纹图谱的研究，建立该复方总黄酮提取物的指纹图谱，标定了23个共有峰，方法重现性和稳定性良好，为复方脑得生总黄酮提取物建立与国际接轨的质量评价方法奠定了基础。采用现代药理学方法，对不同有效部位组方进行了抗血栓活性筛选研究，结果红花、葛根和山楂三味中药混合提取纯化得到的有效部位组方较三味中药分别提取纯化得到的有效部位组方的药效强，并与原制剂脑得生片其抗血栓活性一致，为传统中药复方制剂脑得生片研制为中药有效部位组方的创新中药品种脑得生软胶囊提供了参考依据。采用现代分析方法，对脑得生软胶囊中三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁、葛根素和羟基红花黄色素A等五种主要活性成分进行了含量分析研究，分别建立了应用HPLC-ELSD法测定三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁含量和应用HPLC-UV法测定葛根素和羟基红花黄色素A含量的方法，方法灵敏度高、重现性好、可靠性强。采用HPCE法对脑得生的指纹图谱进行了研究，标定了34个共有峰，建立了图谱清晰、分离度好、可靠性强的指纹图谱测定方法；并对不同厂家生产的脑得生片进行了指纹图谱分析比较。从而为建立该制剂与国际接轨的质量评价方法奠定了基础。采用两制剂双周期交叉试验设计，以兔为试验动物，分别测定灌胃脑得生片和脑得生软胶囊后不同时间血液中主要活性成分葛根素的含量，对脑得生片和脑得生软胶囊进行了药动学和初步生物等效性研究，结果表明脑得生片与脑得生软胶囊在生物上等效，从而为其二次开发提供了理论依据。对脑得生片和脑得生软胶囊的血清药化学进行了初步研究，为阐明其药效物质奠定了一定的基础。更多还原

【Abstract】 The prescription of Naodesheng pian（wan）which is composed ofRadix et Rhizoma Notoginseng,Rhizoma Chuanxiong,Flos Carthami,Radix Puerariae lobatae and Fructus Crataegi comes from ChinesePharmacopoeia 2005 edition.The major actions of this recipeinclude promoting blood circulation to remove blood stasis anddispersing stagnation to remove obstruction in meridiens andcollateralls.It is indicated for sequelae of apoplexy anddizziness due to stagnat blood in in meridians and collaterallsIt is applicable to cerebral arteriosclerosis,stoke and sequelaeof cerebral hemorrhage.It has good effects testified by theclinical applications.Based on pharmacological research,thethesis studies on the antithrombus material basis of traditionalchinese medicinal preparation Naodesheng,including extraction andpurification process,chemical constituents,content analysis,fingerprint,bioequivalence,serum pharmacochemistry etc,aims toprovide bases of the secondary development for the Naodeshengpreparation.Supercritical fluid CO₂ extraction technology was used toextract the essential oil from Ligsticum chuanxiong by orthogonal design,the technological parameters ascertained by the experimentwere proved to be stable and reliable.The antithrombotic functionscreening study on the different extracts from Ligsticum chuanxiongwas carried out.The results show that the effect of oil that reducedsignificantly thrombus wet weight is superior to total extracts,indicate the production process is feasible.Using GC-MS method,35 chemical constituents were identified.The chromatographicfingerprint of supercritical fluid CO₂ extract of Ligsticumchuanxiongwas developed by GC,26 peaks were identified by GC-MS.The method has good reproducibility and stability.The researchshould provide a good foundation of quality evaluation criteria inline with international standards for Ligsticum chuanxiong and itsextract.The method of solvent and the purification technology ofmacroporous adsorption resin were used to study the extraction andpurification process about the effective part of compoundNaodesheng.According to the elution rate and the purity of theproducts,the preparation performance of total flavonoids inNaodesheng by D-101 macroporous resin was investigated,and thepurification technological parameters optimized by the orthogonaltest and single-factor test were proved to be stable and reliable,these can make transition to industrialization.The contents ofpuerarin and hydroxysafflor yellow A were determined by HPLC,andthe total flavonoids with colorimetry.The analysis method waswell repeated and good credibility.The research can effectivelycontrol the quality of total flavonoids extract of Naodesheng.Thefingerprint of the total flavonoids extract of Naodesheng wasstudied by HPLC,with result that 23 peaks were common respectively.The method is stable,well repeated and may offerscientific basis for the quality evaluation of total flavonoidsextract of Naodesheng.The antithrombus pharmacology trial is apply to screen theeffective fraction of Naodesheng from different extractionprocesses.The results show that the preparation of mixedextraction（Flos Carthami Radix Puerariae lobatae and FructusCrataegi）is same effect of antithrombus as the originalpreparation,and superior to that of separate extraction.Theresearch should provide a reference for developing Naodesheng softcapsules made up of active fractions from Naodesheng tablets.The modern analysis methods were carried out to study on thequality standard of Naodesheng soft capsules.The contents ofnotoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 weredetermined by HPLC-ELSD,and puerarin and hydroxysafflor yellow Aby HPLC--UV.The methods are sensitive,reproducible and reliable.The fingerprint of Naodesheng soft capsules was developed byHPCE.Through analyzing the fingerprint of all samples,34 peakswere assigned common peaks in the spectrum of 10 batches.The methodis stable,well repeated.Besides Naodesheng soft capsules,3batches Naodesheng tablets from the different manufacturers alsohas been studied,so that we could find out their commonness anddifference.In short,this research not only elucidate therelationship between different preparation Naodesheng,but alsomay lay the foundation for establishing a international qualitystandard.Naodesheng soft capsules and reference tablets were given to6 rabbits in a randomized crossover design.Puerarin concentrations in plasma were determined by HPLC.The pharmacokinetic parametersand relative bioavailability were calculated with PKSolver 2.0program to evaluate the bioequivalence of the two preparatons.Thestatistical analysis of the results shows that the two preparatonsare bioequivalent.we researched on the serum pharmacochemistry ofNaodesheng soft capsules and Naodesheng tablets,in order to provideguidance for clarifying the material basis.更多还原