菝葜抗炎物质基础及其资源品质研究

【作者】 晏绿金；

【导师】 刘焱文；

【作者基本信息】 湖北中医学院 ， 中医基础理论， 2008， 博士

【摘要】 菝葜(Smilax china L.)又名金刚藤,为百合科植物,药用其根茎。菝葜为少常用中药,收载于《中国药典》2005年版一部,具有祛风利湿,解毒散瘀的功效;现代药理研究表明,菝葜提取物具有抗炎、抗肿瘤及调节免疫功能等作用;以菝葜为原料制成的金刚藤胶囊、金刚藤糖浆等中成药广泛用于临床治疗妇科炎症疾患,疗效显著。本论文在药理试验跟踪下,对菝葜抗炎物质基础及其资源品质进行了较系统研究。采用学科交叉的研究方法,系统筛选了菝葜抗炎药效物质,确定了乙酸乙酯萃取物和正丁醇萃取物为抗炎有效部位。化学成分分析表明,前者主要为黄酮类成分,后者主要为皂苷类成分。采用色谱分离法和波谱鉴定法等现代科学技术,从菝葜药材抗炎有效部位中分离和鉴定了14种化合物,分别为没食子酸(Gallic acid,Ⅰ)、咖啡酸(Caffeic acid,Ⅱ)、原儿茶酸(Protocatechuic acid,Ⅲ)、3,5,4’-三羟基芪(3,5,4’-trihydroxystilbene,Ⅳ)、3,5,2’,4’-四羟基芪(3,5,2’,4’-tetrahydroxystilbene,Ⅴ)、山柰酚(Kaempferol,Ⅵ)、花旗松素(Taxifolin,Ⅶ)、槲皮素(Quercetin,Ⅷ)、胡萝卜苷(β-sitosterol-D-glucoside,Ⅸ)、落新妇苷(Astibin,X)、槲皮素4’-0-β-D-葡萄糖苷(Quercetin-4’-0-β-D-glucoside,Ⅺ)、二氢山柰酚3-0-α-L-鼠李糖苷(黄杞苷,Dihydrokaempferol-3-0-α-L-rha,Ⅻ)、槲皮素3-0-α-L-鼠李糖苷(Quercetin-3-0-α-L-rha,ⅩⅢ)、薯蓣皂苷(dioscinⅩⅣ)。其中化合物Ⅰ、Ⅲ、Ⅶ、Ⅹ为首次从该植物中发现。这对于诠释菝葜抗炎药效物质基础具有重要意义。通过检测大鼠炎足PGE2含量,以及运用PGE2放射免疫法测定菝葜活性部位对大鼠血清血栓素B2(TxB2)及6-酮-前列腺素F1a(6-keto-PGF1a)、IL-1、IL-8、TNF水平的影响,对菝葜抗炎药效物质的作用机理进行探讨。研究结果表明,菝葜抗炎活性物质能显著抑制多种组织源性炎症介质,从而为以菝葜药材为原料生产的中成药制剂的临床疗效提供了科学依据。在菝葜抗炎药效物质研究的基础上,通过中药现代化共性关键技术从菝葜药材中提取纯化抗炎有效部位,即中药标准提取物。采用紫外分光光度法对菝葜抗炎标准提取物中总黄酮和总皂苷进行了含量分析研究,采用高效液相色谱法(HPLC)对菝葜标准提取物的主要有效成分槲皮素进行了含量分析研究,采用HPLC-UV法和HPLC-ELSD法测定了菝葜提取物的指纹图谱,均建立了可靠的分析方法,符合《中国药典》和国家食品药品监督管理局颁布的技术要求。从而为菝葜标准提取物构建了与国际接轨的质量评价体系,也为以菝葜药材作原料生产的中成药二次开发成创新中药品种奠定了坚实基础。为掌握菝葜药材在我国的分布情况,蕴藏量及其生态环境,选择了湖北、湖南、江西、安徽、四川、广东、广西、福建、浙江、重庆等10个菝葜药用植物资源较丰富的省区进行植物资源调查,对其生长的地形地貌,海拔高度、土壤性质等生态环境进行了考察,对其分布情况进行了分析;同时,对不同产地菝葜药材中主要有效成分进行了含量分析比较研究,达到了预期研究目标。这对合理开发利用菝葜药用植物资源,从源头保证以菝葜药材为原料生产的中成药制剂的质量,具有十分重要的意义;从而为完善菝葜药材规范化种植提供了科学依据,并为菝葜药用植物可持续性开发利用奠定了良好的基础。更多还原

【Abstract】 Rhizoma smilaci.popularly known in China as "JinGangTeng",belongs to the Liliaceae family Smilax china L.It is commonly used Chinese medicine,which was collected in "the Chinese Pharmacopoeia"in 2005 version.It has various pharmacological effects including relieve sickness wet and disintoxication and so on.Recent pharmacological study has proved it has anti-tumor activity,anti-inflammatory activity and regulation of the immunity activity and so on.The traditional Chinese medicine preparation of "JinGangTeng capsule "and "JinGangTeng syrup",which are made from the main ingredient of Rhizoma smilaci,have good therapeutic efficacy against variouse gynecological inflammation diseases.Under the pharmacological study,this paper is a systematic research carried on the actively anti-inflammatory substances,plant resources and quality of Rhizoma smilaci..Screen the actively anti-inflammatory constituent from the extraction of Rhizoma smilaci.by interdiscipline.The results showed that both ether acetoacetate fraction and n-Butanol fraction have significantly anti-inflammatory activity.The former is the main in gredient of flavones and the latter saponins.14 compounds were extracted and isolated from active fractions by chromatography and spectroscopy.Their structures have been determined as Gallic acid①,Caffeic acid②,Protocatechuicacid③,3,5,4’-trihydroxystibene④,3,5,2’,4’ -tetrahydroxystilbene(Ⅴ),Kaempferol(Ⅵ),Taxifolin(Ⅶ),Quercetin(Ⅷ),β-sitosterol-D-gluco-side (Ⅸ),Astibin(Ⅹ),Quercetin-4’-O-β-D-glucoside(Ⅺ),Dihydrokaempferol-3-O-a-L-rha(Ⅻ), Quercetin-3-O-a-L-rha(Ⅺ),dioscin(ⅩⅣ).CompoundⅠ,Ⅲ,Ⅶ,X were found in Rhizoma smilaci.for the first time.It is of great significance for Rhizomasmilaci to clarify its materially anti-inflammatory foundation.The mechanism of actively anti-inflammatory substance had been discussed by detecting the content of PGE2 about rat’s paws and determining the lever infection of B2(TxB2), F1a(6-keto-PGF1a),IL-1,IL-8,TNF about rat’s blood serum by radioimmunoassay.The results showed that the actively anti-inflammatory substances of Rhizoma smilaci.have good inhibition on various inflammatory mechanics.So,it provides scientific evidence of clinical effects about the traditional Chinese medicine preparation made from the main ingredient of Rhizoma smilaci..Based on the research of its materially anti-inflammatory foundation,we extracted and purified anti-inflammatory avtivity fraction,which is Chinese herb standard extract,by Chinese traditional medicine modem general character essential technology.The contents of flavones and saponins in standard extract of Rhizoma smilaci.were researched by UV. The content of Quercetin was researched by HPLC,too.Then,the Fingerprints of the standard extract were established by HPLC-UV and HPLC-ELSD.All of above researches are responsible approach,which correspond with the technical requirements of the Chinese Pharmacopoeia(2005)and the State Food and Drug Administration.So,it builded up the quality assessment system geared to the international standards.At the same time,it laid on the hard foundation for the new creation of traditional Chinese medicine preparation made from the main ingredient of Rhizoma smilaci..In order to grasp the information about hizoma smilaci.’s distributed characteristic, reserves and ecological environment.we have investigated the resources of Rhizoma smilaci. in more than 10 provinces such as Hubei,Hunan,Jiangxi,An’hui,Sichuan,Guangdong, Guangxi,Fujian,Zhejiang,Chongqing;studed it’s ecological environment information about terrain,altitude.the nature of soil;analyzed it’s distributed characteristic;also compared to analysis of content in Rhizoma smilaci.from different areas.All expected objectives have been obtained.Thus,it is of great significance to utilize properly resources of Rhizoma smilaci.and control the quality of products made from the main ingredient of Rhizoma smilaci..Meanwhile,the paper provides scientific evidence of perfecting GAP and lays on the hard foundation for the sustainable development and utilization of Rhizoma smilaci..更多还原