【作者】 张存莉；

【导师】 朱玮；

【作者基本信息】 西北农林科技大学 ， 植物学， 2007， 博士

【摘要】 黑刺菝葜（Smilax scobinicaulis C.H. Wright）是菝葜科菝葜属植物,其根茎具有除风湿、活血解毒、镇惊息风、治疗肠炎、疮疖、癌肿等功效。杨梅（Myrica rubra Sieb et Zucc）是杨梅科杨梅属植物,其根可止血理气,治疗胆囊炎、牙周炎和疝气等病。然而,对黑刺菝葜化学成分及生物活性的系统研究和杨梅根化学成分研究未见报道。本论文对黑刺菝葜次生代谢产物的分布规律、黑刺菝葜提取物的抑菌活性、黑刺菝葜和杨梅根化学成分的分离和结构及黑刺菝葜甾体皂苷的抑菌活性进行了研究,从而为菝葜属植物和杨梅属植物的化学分类学研究及其药理作用的阐明提供了一定的理论依据。主要研究结果如下:1.采用化学成分系统预示和分光光度法首次对黑刺菝葜不同器官次生代谢产物的分布规律和主要成分含量进行了研究,结果表明黑刺菝葜次生代谢产物在植株不同器官中的种类及含量有差异。除果肉和果实中不含甾体皂苷、香豆素和内酯类化合物,只有果核中含油脂,果肉中含有强心苷类化合物外,黑刺菝葜各器官都含有糖类、酚类、鞣质、氨基酸、蛋白质、生物碱、皂苷、黄酮、蒽醌、香豆素、甾体、内酯等化学成分。在不同器官中,黄酮类化合物含量高低顺序为:须根>果肉>块根>叶>茎>根茎>茎卷须>果核,甾体皂苷类化合物含量的高低顺序为:叶>茎>卷须>块根>根茎>须根。2.对黑刺菝葜根、茎、叶醇提物的石油醚、乙酸乙酯、正丁醇和萃取后剩余水相,以及根萃取后剩余水相大孔吸附树脂洗脱物共17种提取物进行了室内抑菌活性测试。结果表明:黑刺菝葜不同器官和同一器官极性不同的萃取物对供试菌抑制活性不同,且普遍对细菌有抑制作用,对真菌几乎不显示抑制作用,其中茎乙酸乙酯提取物对供试细菌:金黄色葡萄球菌、枯草芽孢杆菌、蜡状芽孢杆菌、大肠杆菌、巨大芽孢杆菌和普通变形杆菌的抑菌作用最强,其MIC分别为0.8 mg·mL^-1、0.6mg·mL^-1、1.2 mg·mL^-1、1.0 mg·mL^-1、1.0 mg·mL^-1和1.0 mg·mL^-1。根剩余水相大孔吸附树脂醇洗脱物中只有100%乙醇洗脱物表现出极强的广谱性抗菌活性,对金黄色葡萄球菌、枯草芽孢杆菌、蜡状芽孢杆菌、大肠杆菌、普通变形杆菌、巨大芽孢杆菌、黄曲霉、黑曲霉、康氏木霉、青霉和绿色木霉的MIC分别为2.0 mg·mL^-1、1.0 mg·mL^-1、1.2 mg·mL^-1、1.0 mg·mL^-1、1.2 mg·mL^-1、1.5 mg·mL^-1、0.5 mg·mL^-1、0.5 mg·mL^-1、0.8 mg·mL^-1、0.8 mg·mL^-1和0.3 mg·mL^-1;此外,80%乙醇的洗脱物对绿木霉、大肠杆菌显示出极强的抑制活性,其MIC分别0.2 mg·mL^-1和0.5 mg·mL^-1,其它洗脱物对供试菌几乎没有抑制作用。3.从黑刺菝葜根茎中分离得到20个甾体皂苷,鉴定了13个化合物的结构,Scifinder检索结果表明其中有7个新化合物,6个已知化合物。新化合物有:（25S）-?5-螺甾烯-3β,27-二醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（1）,（25S）-螺甾-3β,11,27-三醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（3）,（25S）-螺甾-3β,27-二醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（4）,（25S）-螺甾-3β,27-二醇-3-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（5）,（25R）-螺甾-6β-醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（7）,（25S）-?5-螺甾烯-3β-醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（8）,（25S）-螺甾-3β,17α,27-三醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（9）。已知化合物有:异娜草皂苷元-3-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（2）,（25R）-替高皂苷元-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（6）, （25S）- ?5-螺甾烯-3β,17α,27-三醇-3-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（10）,（25S）- ?5-螺甾烯-3β,17α,27-三醇-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（11）,拉克索皂苷元-3-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（12）,拉克索皂苷元-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（13）。4.从黑刺菝葜根醇提物的正丁醇萃取物中筛选到一批抗细菌甾体皂苷活性物质,其中对普通变形杆菌抑制作用较强的是:大孔吸附树脂50%乙醇洗脱物（R-5）、70%乙醇洗脱物（R-7）、80%乙醇洗脱物（R-8）和90%乙醇洗脱物（R-10）,对大肠杆菌抑制作用较强的是:60%乙醇洗脱物（R-6）、R-8、R-9和R-10;对巨大芽孢杆菌抑制作用较强的是:R-7、R-8、R-9及拉克索皂苷元-3-O-[α-L-吡喃阿拉伯糖基（1→6）]-β-D-吡喃葡萄糖苷（皂苷1）和拉克索皂苷元-3-O-[β-D-葡萄吡喃糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（皂苷2）;对蜡状芽孢杆菌和枯草芽孢杆菌抑制作用较强的是:R-7、R-8和R-9。从黑刺菝葜根醇提物的正丁醇萃取物中筛选到一批抗真菌活性物质,除正丁醇萃取物以外,所有皂苷样品对绿色木霉都有很强的抑制作用;对黑曲霉抑制作用较强的是R-7、R-8、皂苷1和（25S）-3β,27-二羟基-5α-螺甾-6-酮-3-O-[β-D-吡喃葡萄糖基-（1→4）-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（皂苷3）;对康氏木霉抑制作用较强的是R-9、皂苷1～3和（25S）- 5烯-螺甾-3β,17α,27-三醇-3-O-β-D-吡喃葡萄糖-（1→4）-[α-L-吡喃阿拉伯糖（1→6）]-β-D-吡喃葡萄糖苷（皂苷4）;对青霉抑制作用较强的是R-7、皂苷1、皂苷2和皂苷4;对黄曲霉抑制作用较强的是R-7、皂苷1、皂苷3和皂苷4。5.从黑刺菝葜根醇提物的乙酸乙酯萃取物中首次分离得到4个化合物,其结构鉴定为:β-谷甾醇（1）、拉克索皂苷元（2）、槲皮素（3）和胡萝卜苷（4）。其中拉克索皂苷元为首次从该植物中分离得到。6.从黑刺菝葜茎中首次分离得到5个化合物,其结构鉴定为:反式白藜芦醇（1）、槲皮素-3-O-β-D-葡萄糖苷（2）、木犀草素-7-O-β-D-葡萄糖苷（3）、拉克索皂苷元-3-O-[α-L-吡喃阿拉伯糖基-（1→6）]-β-D-吡喃葡萄糖苷（4）、拉克索皂苷元-3-O-{β-D-吡喃葡萄糖基-（1→4）-[α-L-吡喃阿拉伯糖基-（1→6）]}-β-D-吡喃葡萄糖苷（5）。其中化合物1～3为首次从该植物中得到。7.从杨梅根中首次分离得到7个化合物,其中化合物1～5为环型二芳基庚酮类及其衍生物类,分别为:杨梅醇（1）、杨梅醇-5-O-β-D-葡萄糖苷（2）、8-羟基杨梅酮（3）、杨梅酮-5-O-β-D-葡萄糖苷（4）和porson（5）,另外两种化合物是胡萝卜苷（6）和β-谷甾醇（7）。化合物3是新化合物,化合物5～7为首次从杨梅中得到。更多还原

【Abstract】 Smilax scobinicaulis C.H. Wright belongs to family of Smilacaceae. Its rootstalk has been used in folk medicine as antirheumatic disease, move blood stasis and clear toxins, anticonvulsant, curing enteritis and cancer etc. Myrica rubra Sieb et Zucc belongs to family of Myricaceae. Its root can be used as hemostatic and to cure cholecystitis, pericementoclasia and colic etc. However, the chemical constituents and biological activities of the root from Smilax scobinicaulis and Myrica rubra have not been reported. To further exploit and utilize these plant resources and provide a theoretical basis on taxonomy of chemical and clarify the role of pharmacological for Smilax and Myrica plant, this dissertation were carried out a systematic research on specific distribution property of chemical constituents from Smilax Scobinicaulis, the antimicrobial activity of the extract of Smilax scobinicaulis, The separation and structure of the chemical composition from Smilax scobinicanlis and the root of Myrica rubra. The result showed as follows:The distribution property of chemical constituents from Smilax scobinicaulis was studied through the protested systematically and UV method. The result indicated that organs almost have sugar, phenol, digallic acid, aminoacid, protein, alkaloid,saponin, anthoxanthin, enthraqu -inone, coumarin,sterol and lactones etc.except saponin, oumarin and lactones in fruit , fat only in kernel and cardiac glycoside only in pulp. The totalcontent of flavonoids:fibrous root> pulp> root tuber> leaf>stem > rhizome> tendril> kernel. The total content of saponin:leaf > stem > tendril>root tuber> rhizome> fibrous root.Filter-paper diffusion method was adopted in lab to test the antimicrobial activities of seventeen extracts, which concluds twelve solvent extracts and five eluates obtained with macroreticular resin at different ethanol concentrations from the root, stem and leaf of Smilax scobinicaulis. The result revealed that different organs and different extract of the same organ of Smilax scobinicaulis have different inhibiting activity on animalcule, generally inhibit bacteria and almost no effect on fungus. The ethyl acetate extract from stem have strongest inhibitory effect on the tested Staphyloccocus aureus, Bacillus subtilus, B. cereus, E. coli, B. megaterium and Proteus vulgaris. Its MIC respectively is 0.8mg·mL^-1, 0.6mg·mL^-1, 1.2 mg·mL^-1, 1.0mg·mL^-1, 1.0mg·mL^-1 and 1.0mg·mL^-1. The eluate obtained by macroporous resin with ethanol at a concentration of 100% showed a very strong broadband antimicrobial activity. Its MIC on S. aureus, B. subtilus, B. cereus, E. coli, P. vulgaris, B. megaterium, Aspergillus flavus, Aspergillus niger, Trichoderma Koningii, Penicillium sp. and Trichoderma viride respectively is 2.0mg·mL^-1, 1.0mg·mL^-1, 1.2mg·mL^-1, 1.0mg·mL^-1,1.2mg·mL^-1, 1.5mg·mL^-1,0.5mg·mL^-1, 0.5 mg·mL^-1, 0.8mg·mL^-1, 0.8mg·mL^-1and 0.3 mg·mL^-1. Moreover, the eluate obtained by macroporous resin with ethanol at a concentration of 80% showed very strong inhibitory activity on Trichoderma viride and E. coli. Its MIC respectively is 0.2 mg·mL^-1 and 0.5 mg·mL^-1. Else eluate has no inhibitory activity on animalcule.Thirteen steroidal saponins were separated from the root of Smilax scobinicaulis and their structures were identified. The result showed that seven steroidal saponins are new compounds and six steroidal saponins are known compounds by scifinder search. The new compound respectively is （25S）-spirostan-5-ene-3β,27-diol（isonarthogenin）3-O-{β-D- glucopyrnosyl -（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （1）,（25S）-spir -ostan-3β,11,27-triol-3-O-{β-D-Glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （3）, （25S）- spirostan-3β, 27-diol-3-O-{β-D-Glucopyrnosyl-（1→4）-[α-L- Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （4）, （25S）-spirostan-3β,27-diol-3-O-[α-L- Arabinopyranosyl-（1→6）]-β-D-glucopyranoside （5）,（25R）-spirostan-6β-ol-3-O-{β-D- glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （7）, （25S）- spirostan-5-en-3β-ol-3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （8）, （25S）-spirostan-3β,17α,27-triol-3-O-{β-D-glucopyrnosyl-（1→4）-[α-L- Arabinopyranosyl-（1→6）]}-β-D-Glucopyrano-side （9）.The known compound respectively is （25S）- spirostan-5-ene- 3β,27-dihydroxy-3-O-[α-L -Arabinopyranosyl-（1→6）]-β-D-glucopyranoside （2）, （25R）-5α-spirostan-3β-ol （tigogenin） 3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （6）,（25S）-spirostan-5-en-3β,17α,27-triol-3-O-[α-L-Arabinopyranosyl-（1→6）]-β-D-glucopyranoside （10）,（25S）-spirostan-5-en-3β,17α,27-triol-3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-arabinopy arabinopyranosyl-（1→6）]}-β-D-Glucopyranosid （11）,（25R）-3β-hydroxy-5α-spirostan-6-one（laxogenin）3-O-[α-L-Arabinopyranosyl-（1→6）]-β-D-glucopyranoside （12）,laxogenin-3 -O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside （13）. A group of anti-bacterial steroidal saponins from n-butanol extraction of the root from Smilax scobinicaulis were screened. The eluates obtained by macroporous resin with ethanol at a concentration of 50%（ R-5）, 70%（R-7）, 80%（R-8）and 90%（R-9）have stronger inhibitory activity on P. vuigaris; The eluates obtained by macroporous resin with ethanol at a concentration of 60%（R-6）, R-8, R-9 and R^-10 have stronger inhibitory activity on E. coli.; R-7, R-8, R-9 and Laxogenin-3-O-[α-L-Arabinopyranosyl-（1→6）]-β-D-glucopyranoside（saponin 1）,laxogenin-3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]} -β-D-Glucopyranosid（saponin 2）have stronger inhibitory activity on B. megaterium; R-7、R-8 and R-9 have stronger inhibitory activity on B. cereus and B. subtili.A group of anti-fungi steroidal saponins from n-butanol extraction of the root from Smilax scobinicaulis were also screened. All of steroidal saponins except n-butanol extraction have strong inhibitory activity on T. viride; R-7, R-8, saponin 1 and （25S）-spirostan-3β, 27- diol-6-one -3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinpyranosyl-（1→6）]}-β-D-Glucopyra -noside（saponin 3）have strong inhibitory activity on A. niger; R-9 , saponin1 3 and （25S）-spirostan-5-en-3β,17α,27-triol-3-O-[α-L-Arabinopyranosyl-（1→6）]-β-D-Glucopyranosi-de（saponin 4）have strong inhibitory activity on T. Koningii; R-7, saponin 1, 2 and 4 have strong inhibitory activity on Penicillium sp.; R-7, saponin 1, 3 and 4 have strong inhibitory activity on A. flavus.Four compounds were separated from ethyl acetate extract of the root from Smilax scobinicaulis and their structure were identified asβ-sistostero（l1） , laxogenin（2）, quercetin （3）and daucosterol（4）. Compound 2 was isolated from the roots of the plant for the first time.Five compounds were separated from the stem of Smilax scobinicaulis and theirs tructure were identified as resveratol（1）, querectin-3-O-β-D-glucoqyranoside（2）, luteolin-7-O-β-D-glucoside（3）, laxogenin-3-O-[α-L-Arabinopyranosyl-（1→6）]-β-D-Glucopyranoside（4）, laxogenin-3-O-{β-D-glucopyrnosyl-（1→4）-[α-L-Arabinopyranosyl-（1→6）]}-β-D-Glucopyranoside（5）. Compound 1³ were isolated from the roots of the plant for the first time. Seven compounds were separated from the stem of Smilax scobinicaulis and their structure were identified as myricanol（1）, myricanol-5-O-β-D-glucopyranoside（2）, 8-hydroxy- myricanone（3）, myricanone -5-O-β-D- glucopyranoside（4）, porson（5）, daucosterol（6）,β-sistostero（l7）. Compound 1⁵ were biphenyl-type diarylheptanoid. Compound 3 was a new one. Compound 5⁷ were isolated from the roots of the plant for the first time.更多还原