【作者】 李斌；

【导师】 李顺祥；

【作者基本信息】 湖南中医药大学 ， 方剂学， 2014， 博士

【摘要】 细叶石仙桃(Pholidota cantonensis Rolfe.)为兰科石仙桃属植物,主要分布于我国的云南、广东、广西、贵州、浙江、福建、湖南、江西等省。全草入药,味微甘,性凉,具有清热凉血、滋阴润肺、消肿止痛的功效,民间常用于治疗咽喉肿痛、乳蛾、咳嗽痰喘、肺炎、咳血、跌打损伤和关节炎等。目前对细叶石仙桃成分和药理作用研究较少,为进一步深入研究和开发细叶石仙桃资源,本文采用柱色谱法、高效液相色谱法、制备薄层及重结晶等方法对细叶石仙桃全草80%乙醇提取物的化学成分进行了系统分离,通过1H-NMR、13C-NMR、2D-NMR及MS等光谱方法及理化常数鉴定化合物结构;采用试管稀释法和琼脂扩散法测定了细叶石仙桃不同极性提取物及部分化合物的最低抑菌浓度(MIC)和抑菌圈,采用微量肉汤稀释法测定了部分单体化合物的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。从细叶石仙桃中分离得到31个化合物:BatatasinⅢ(1),Gigantol(2),Thunalbene(3),Orchinol(4),细叶石仙桃菲(5),Ephemeranthoquinone(6),Densiflorol B(7),细叶石仙桃醌(8),丹参酮ⅡA(9),Neo-tanshinlactone(10),丁香醛(11),丁香酸(12),3,5-二甲氧基-4-羟基-苯丙酮(13),紫丁香苷(14),桂皮酸(15),肉桂酸正丁酯(16),E-Ferulic acid docosyl ester(17),丁香脂素(18),TangshennosideⅢ(19),细叶石仙桃苷A(20),细叶石仙桃苷B(21),二十四甲基环阿尔廷醇(22),二十四甲基环阿尔廷酮(23),24-Methylenepollinastanone(24),里白烯(25),麦角甾醇过氧化物(26),β-谷甾酮(27),豆甾醇(28),β-胡萝卜苷(29),β-谷甾醇(30),正二十二醇(31)。包括芪类化合物10个,酚类化合物9个,二萜类苷类化合物2个,三萜类化合物4个,甾体类化合物5个,脂肪醇1个。其中新化合物有4个,包括2个新的芪类化合物和首次从石仙桃属植物中发现的2个新的二萜苷类化合物。化合物9-10,12,14-16,19,23-25,27-28,31为首次从石仙桃属植物中分离得到。细叶石仙桃不同极性提取物对白假丝酵母菌和金黄色葡萄球菌、表皮葡萄球菌、消化链球菌的抑制作用明显。细叶石仙桃提取物对革兰阳性菌和白假丝酵母菌MIC范围为80-120 mg/mL,对革兰阴性菌的MIC范围为100-160mg/mL。化合物1,4,18,19对金黄色葡萄球菌有较好的抑菌效果,MIC均小于25μg/mL;化合物1,3,4,19对绿脓杆菌抑制效果较好,MIC均小于25μg/mL,尤其是化合物3对绿脓杆菌的抑菌效果最明显,其MIC和MBC均为11.67μg/mL;化合物1,3,4,18,19对大肠埃希菌有抑菌效果,MIC与MBC均在50μg/mL左右;化合物1,3,4,19对白假丝酵母菌有抑菌效果,MIC与MBC均在50μg/mL左右。这些化合物的发现和抗菌活性的研究丰富了细叶石仙桃化学成分的内容,揭示了细叶石仙桃的药效物质基础,为进一步系统深入地研究细叶石仙桃的药效物质基础和质量控制等奠定了基础,新化合物的发现丰富了天然产物的内容,为新的生物活性的发现提供了物质基础。更多还原

【Abstract】 Pholidota cantonensis Rolfe.,a medicinal plant,mainly distributed in Yunnan,Guangdong,Guangxi,Guizhou,Zhejiang,Fujian,Hunan,Jiangxi Provinces etc.of China,It is a Chinese folk herbal medicine with a long history of safe use for treating sore throat,tonsillitis,cough phlegm asthma,pneumonia,acute soft-tissue trauma and arthritis,etc.However,as for P.cantonensis,one of the typical species in South China,the relevant chemical and pharmacological studies are limited.In order to further research and development the resources of P.cantonensis,Studies have been conducted on the active components leading to antibacterial effects on the herb.The 80%ethanol extract of the whole plant P.cantonensis was separated by chromatography on silica gel,Sephadex LH-20,MCI gel and pre-HPLC.The structures of compounds from P.cantonensis were elucidated on the basis of the chemical evidence and 1H-NMR,13C-NMR,2D-NMR and MS spectral.The minimum inhibitory concentration(MIC)and the inhibition zone of different extracts from P.cantonensis were measured by tube dilution method and agar diffusion method;the MIC and the minimum bactericidal concentration(MBC)of some compounds were determined by the broth microdilution method.31 compounds were isolated from P.cantonensis:Batatasin Ⅲ(1),Gigantol(2),Thunalbene(3),Orchinol(4),Phocantol(5),Ephemeranthoquinone(6),Densiflorol B(7),Phocantone(8),Tanshinone IIA(9),Neo-tanshinlactone(10),Syringaldehyde(11),Syringate(12),3,5-dimethoxy-4-hydroxy-propiophenone(13),Syringin(14),Cinnamic acid(15),Butylcinnamate(16),E-Ferulic acid docosyl ester(17),Syringaresinol(18),Tangshennoside Ⅲ(19),Phocantoside A(20),Phocantoside B(21),24-methylenecycoartanol(22),24-methylenecycloartone(23),24-Methylenepollinastanone(24),Hop-22(29)-ene(25),Ergosterol peroxide(26),β-sitostenone(27),Stigmasterol(28),Daucosterol(29),β-Sitosterol(30),1-docosanol(31).Including 10 stilbenoid compounds,9 phenolic compounds,2 diterpene glycosides,4 triterpenoids,5 steroids and 1 fatty alcohol.there are four new Compounds:Compound 5,8 are new stilbenoid compounds,Compound 20,21 are new diterpenoids,it is the first time that diterpenoids were isolated from the genus Pholidota,and compounds 9-10,12,14-16,19,23-25,27-28,31 were isolated from P.cantonensis for the first time.The extracts of P.cantonensis had more inhibiting effect on Candida albicans and gram-positive bacteria such as Staphylococcus aureus,Staphylococcus epidermidis and Peptostreptococcus micros than that on gram-negative bacteria.MIC of different extracts of P.cantonensis Rolfe.against Candida albicans and gram-positive bacteria were in the range of 80-120 mg/mL,and that of gram-negative bacteria in the range of 100-160mg/mL.Compounds 1,4,18,19 had good inhibitory effect on Candida albicans,the MIC were less than 25 μg/mL;compounds 1,3,4,19 had good inhibitory effect on Pseudomonas aeruginosa,the MIC were also less than 25μg/mL,especially the compound 3 has showed good bacteriostatic action and bactericidal action,and the MIC and MBC of compound 3 against Pseudomonas aeruginosa were 11.67 μg/mL;Compounds 1,3,4,18,19 showed bacteriostatic action and bactericidal actiont against Escherichia coli,the MIC and MBC were about 50 μg/mL;Compounds 1,3,4,19 showed bacteriostatic action and bactericidal actiont against Canidia albicans too,the MIC and MBC were about 50 μg/mL.The finding of the compounds and the research on the antimicrobial activity enriched the chemical compositions of P.cantonensis,laid the foundation for the further study of the effective substance and the quality control for P.cantonensis,the discovery of new compounds enriched the content of natural product,provided the material basis for new bioactive discovery.更多还原