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题名: 三种茜草科植物化学成分及其抗菌活性研究
作者: 康文艺
学位类别: 博士
答辩日期: 2003
授予单位: 中国科学院昆明植物研究所
授予地点: 中国科学院昆明植物研究所
导师: 郝小江
关键词: 茜草科 ; 化学成分 ; 抗耐药菌株活性
学位专业: 植物学
中文摘要: 本博士期间的研究工作,由两部分组成。上篇论述了三种茜草科植物化学成分的研究,分为三章。从新乌檀属(Neonauclea)的植物无柄新乌檀(N.sesslifolia Merr.);滇丁香属(Luculia)的植物滇丁香(L.pinciana Hook)和帽蕊木属(Mitragyna)的植物帽蕊木(M rotundifolia(Roxb.)Kuntze)中共分离得到了93个化合物,鉴定了其中87个化合物,其中包括9个新化合物。鉴定化合物类型包括三菇、三菇昔、环烯醚昔、幽醇、黄烷醇、黄酮、黄酮昔、香豆素、叫噪类生物碱、芳香化合物等。下篇论述了三种植物抗菌及其它生物活性的研究,分为三章。对三种茜草科植物进行了抗菌活性,尤其是抗耐药菌株活性的筛选。系统地对分离得到的化合物进行了与青霉素协同抗耐药金黄色葡萄球菌活性的筛选研究,结果显示个别化合物具有耐药金黄色葡萄球菌抑制活性;与青霉素配伍,发现了天然产物在抗耐药菌株方面的新用途;利用液体培养对耐药机制进行初步探讨,并寻找到部分化合物与青霉素协同作用时,对耐药金黄色葡萄球菌的抑制率和工IC_(50)此外,对三种植物进行了抗肿瘤活性的筛选,发现无柄新乌檀和滇丁香的提取物具有细胞毒活性,文献未见报道,其结果为这两种植物的潜在开发利用提供了一些科学依据。最后对近年来抗菌药物的发展、耐药菌株的研究进行了综述。目前,抗菌新药的研究与开发,主要是通过合成、半合成方法以及寻找新的微生物菌种,发现新的抗菌药物和抗生素。从植物中,尤其是药用植物中寻找具有抗耐药菌株活性成分的研究,一直是一个不被重视的方向。我们以无柄新乌檀植物提取物具有抗耐药金黄色葡萄球菌为起点,系统地开展了抗耐药菌株活性研究,国内外尚不多见。我国有着丰富的中草药资源,很多中草药的作用是通过抑菌作用表现出来的。因此,从中草药中提取化学成分,筛选具有抗耐药菌株成分的药物,将会丰富中药资源方向的研究内容。一、三种茜草科植物的化学成分研究无柄新乌檀(N.sesslifolia Merr.)为新乌檀属植物,我国有4种,云南分布有新乌檀、无柄新乌檀和具柄新乌檀,在产地作为常用森林用材,未见前人对该属植物化学成分和生理活性的研究。从无柄新乌檀枝干乙醇提取物中共分离鉴定了22个化合物,主要包括三萜、三萜苷、环烯醚苷、香豆素、幽醇和芳香化合物,其中有4个新化合物8,16-18。滇丁香(L,Pinciana Hook)为滇丁香属植物,我国有滇丁香和鸡冠滇丁香2种,分布在广西、云南、贵州和西藏等地,其花、根、果入药,治疗百日咳、慢性支气管炎、肺结核、月经不调、痛经等症,文献中仅见其丹皮酚成分的分离报道。从滇丁香枝干乙醇提取物中,共分离了42个化合物,鉴定了其中的35个,主要包括三萜、三萜苷、黄酮、黄酮苷、环烯醚苷、香豆素、芳香化合物等,其中化合物14,19,20,26为新化合物。帽蕊木(M.rotundifolia(Roxb)Kuntze)为帽蕊木属植物,我国仅有1种,产于云南南部,海拔1000米处的密林中。国外在60-80年代,对该属植物化学成分和药理活性的研究比较活跃,化学成分主要集中在喇噪类生物碱,并对其进行了抗肿瘤、心血管疾病和抗菌活性的研究。我们对帽蕊木茎皮乙醇提取物进行了化学成分的研究,从中分离鉴定了2S个化合物,主要为三菇、三菇昔、黄烷醇、叫噪类生物碱、幽醇和芳香化合物等,其中化合物1为一新的三菇昔。二、三种植物抗菌及其它生物活性研究将从三种植物中分离得到的部分化合物进行了抗菌试验,发现无柄新乌檀中的化合物4对金黄色葡萄球菌的最小抑制浓度(MIC25μg/片);滇丁香中化合物10对金黄色葡萄球菌具有抑制和杀灭作用(MIC 0.5mg/ml;MBC10mg/ml):2对白色念珠菌具有抑制和杀灭作用(MIClmg/ml; MBC5mg/m1)。将三种植物中分离得到的42化合物,进行了固体平板耐药金黄色葡萄球菌活性的研究,发现其中10个化合物具有抗耐药金黄色葡萄球菌的活性,其化合物的结构类型包括1个蒽醒类、1个芳香化合物、1个黄烷醇、7个三枯及其苷。在上述抗菌活性的基础上,我们又进行了与青霉素和甲氧节吮配伍的协同增效试验,并用市场上的产品新达宝和艾可儿做为评价指标,发现帽蕊木中分离得到的化合物21(MIC 0.57μg/片),滇丁香中分离得到的化合物23(MIC 0.565μg/片)与青霉素配伍比列与市场产品相近似。我们在固体平板培养的基础上,进行了液体培养,建立了该菌株的生长曲线,并对部分化合物与青霉素的协同作用,进行了抑制率的研究,确定了滇丁香中分离的化合物10与青霉素协同作用下的IC50=7.39μg/ml(12 hrs)和12μg/1(24 hrs);化合物20工C50=14.27 pg/ml(12 hrs)和50.18μg/ml(24hrs);帽蕊木中分离的化合物21与青霉素协同作用下IC50=45.99μg/m1(12hrs)和58.53μg/ml(24 hrs)。并初步在液体培养中,探讨该菌株产生耐药的机制。无柄新乌檀、滇丁香和帽蕊木的乙醇提取物进行了P388小鼠白血病与A549人肺腺癌细胞株抑制活性的分析,显示三种植物的乙醇提取物对这两个细胞株均具有抑制活性。在此基础上,对前两个植物分离得到的部分化合物进行人HL-60白血病与Hel。宫颈癌细胞株抑制活性的分析,发现滇丁香植物中分离得到的化合物19在100μg/ml时,对HL-60细胞株的抑制率57.97%,对Hela细胞株抑制率32.08%。最后对抗菌药物的发展和耐药菌株的研究进行了论述。
英文摘要: The thesis mainly contains two parts, in which the chemical constituents of three plants, Neonaucka sesslifolia Merr., Luculia pinciana Hook., and Mitragyna rotundifolia (Roxb.)Kuntze.. have been investigated. Ninety and three including nine new compounds were isolated and identified. Theses compounds including triterpenoids and saponins, iridoid glycoside, sterol, flavanes, flavones and glycosides, coumarins, indole alkaloids and aromatic derivatives. 1. Chemical Constituents from Neonauclea sessilifolia Merr The plant of N sessilifolia (Rubiaceae), is distributed abundantly in xishuangbanna of Yunnan province as timber. There were no chemical constituents and bioactivity reported for this genus, previously. Twenty and two compounds were isolated from the stems of N. sessilifolia., mainly including triterpenoids and saponins, iridoid glycoside, coumarins, sterol and aromatic derivatives. They were Paeonol (1), 3, 4-dihydroxybenzoic acid (2), Scopletin (3), Anthraquinones chrysophanol (4), 4H-l-benzopyran-4-one (5), 5,7-dihydroxy-2-methyl (6), P-Sitosterol (7), stigmasterol glacoside (8), Quinovic acid-3-O-a-L-rhamnopyranosyl (l→4)-P-D-glucopyranoside (9), Quinovic acid-3-O-6-deoxy-P-D-glucopyranoside (10), Quinovilic acid-3-O-P-D-Glucopyranoside (11), cincholic acid-3-0-6-Deoxy-P-D-Glucopyranoside(12), Quinovic acid-3-O-6-deoxy-P- glucopyraosyl(28→l)-p-D- glucopyranosyl ester (13), 3-Hydroxy-12-oleane-(28→l)-P-D-glucopyranosyl ester (14), Ursen acid-3-O-p-D-glucopyranoside (15), 3,15,21,23-tetrahydroxy-olean-12-en-28-oic acid (16), 3, 6, 21, 23-terahydroxy-olean-12-ene-28-oic acid (17), 3, 7, 21, 23-Tetrahydroxy-olean-12-ene-28-oic acid (18), Quinovic acid-3-0-p-D-glucopyranosyl(1→3)-6-deoxy-p-glucopyranoside (19), Oleanic acid-3-O-P-D- xylopyranosyl-(l→2)-β-D- glucopyranoside-28-O-P-D-glucopyranosyl ester (20), Iridoid loganin (21), 7-methoxy-gentiopicro side(22), respectively. Among them, compounds 8,16,17 and 18 were new. Luculia pinciana (Rubiaceae) is a kind of medicinal plant widely distributed in Yunnan and Guangxi province of China. As a frequently used drug for tracheitis, tuberculosis, and rheumatic diseases in traditional Chinese medicine, it can "relieve pain, dredge all the channels and vessels, promote blood circulation". The first investigated constituents of the plant matter were paenol and volatile oil, and the pharmacological experiments proved that the ethanol extract had antibiotic activities. In the course of phytochemical studies on this plant by bio-guided fraction, thirty and five compounds were isolated from the extracts of the stems of L. pinciana., mainly including triterpnoids and saponins, flavones and glycosides, iridoid glycoside, coumarins and aromatic derivatives. They were Scopletin (1), 2, 4 -Dihydroxyacetophenone (2), 5-Methoxy-8- hydroxycoumarin (3), Daphnetin (4), 4 -Hydroxyphenyl methyl ketone (5), 3-Hydroxy-4-methylloxy-benzeic acid (6), 5-Hydroxyl-2-methyl-benzeoic acid (7), 3-(4 -Hydroxyphenyl)-2-propenoic acid (8), Dauricine (9), Ursolic acid (10), Oleanolic acid (11), Vogeloside (12), epi-Vogeloside (13), 4-[4 -O-(2 ,3 ,5 ,6 -Tetrahydroxyphenyl)-p-glucopyranoside]-l-butene (14), Glucose-1- [3-(3 -methoxy-4 -hydro xy) phenyl]-2-cis propenoate (15), Loganoside (16), Loganin (17), 5, 7, 4-tetrhydroxy-3-methoxyflavone (18), la, 30, 24-trihydroxy-urs-12-en-28-ic acid (19), Quinovic acid-28-O-p-D-glucopyranosyl ester (20), Quriovic acid-3-O-a-L-rhamnopyranoside-28-O-p-D-glucopyranosyl ester(21), Quinovic acid-3-O-P-D-glucopyrano side (22), Qunovic acid-3-O-p-D-glucopyranoside-28-O-p-D-glucopyranosyl ester (23), Quinovic acid-3-O-P-6-deoxy-D-glucopyranoside-28-O-p-D-glucopyranosyl ester (24), Quinovic acid 3-O-P-6-deoxy-D-glucopyranoside (25), Cincholic acid-3-O-p-D-xylopyranoside, 28-O-P-D-glucopyranosyl ester (26), Cincholic acid-3-O-p-D-glucopyranoside, 28-O-p-D-glucopyranosyl ester (27), Cincholic acid-3-O-P-D-glucopyranoside (28), Kaempferol 3-O-a-L-rhamnoside(29), Kaempferol 3-O-rutinoside (30), Kaempferol 3-O-p-D-glucopyranoside-7-O-a-L-rhamnoside (31), Kaempferol 3-0-rutinoside-7-O-a-L-rhamnoside (32), Kaempferol 3-O-a-L-rhamnoside-7-O-(3-D-gluco pyranosyl(l→2)-p-D-glucopyranoside (33), D-Glucopy ranosyl(l-~6)-D-Glucose (34), and Stearic acid (35), respectively. Among them, compounds 14,19, 20,26 were new. Twenty and eight compounds were isolated from the extracts of the bark of M rotundifolia, mainly including triterpenoids and saponins, iridoid glycoside, sterol, flavanes, coumarins, amides, indole alkaloids and aromatic derivatives, they were Quinovic acid-3-0-P-D-6-Deoxy-glucopyranoside, 28-O-p-D-glucopyranosyl ester (1), Quinovic acid-27-O-a-L-Rhamnopyranosyl ester (2), Quinovic acid-3-O-a-L-Rhamnopyranoside (3), Qunovic acid-27-O-P-D-glucopy ranosyl ester (4), Quovic acid-3-O-P-D-6-Deoxy-glucopyranoside (5), Quovic acid-3-O-p-D-glucopyranoside (6), Cincholic acid-P-D-6-Deoxy-glucopyranoside (7), Cincholic acid-28-O-P-D-glucopyranosyl ester (8), Cincholic acid-3-O-P-D- glucopyranoside (9), Dauricinec (10), 3, 4-dihydroxybenzoic acid (11), p-sitosterolol (12), Scopleton (13), 3, 4, 5-trimethoxyphenol-1-glucopyranoside (14), Taraxerol (15), 4-hydroxy, 3-methyloxy-benzoic acid (16), 3-hydroxy, 4-methyloxy-benzoic acid (17), Caffeic acid (18), 4 -methylepiganocotechm 5-O-p-D-glucopyran oside (19), 3, 4, 5, 5, 7-pentahydroxyflavan (20), 3, 3', 5, 5, 7-pentahydroxy flavan (21), 3, 3*, 4', 5, 7-pentahydroxyflavan (22), 3', 4, 4', 5, 7-pentahydroxyflavan (23), l,l-dimetheyl-2-acetl-diethyl ether (24), Gambirine (25), Gambireine (26), Arachidic acid (27), and acetic acid (28), respectively. Among them, compound 1 was new, and compounds 5,6, 8-13, 16 ,17 were also isolated from N. sessilifolia and L. pinciane, 2. Antibiotic bacteria biological assay of three plants The activity to inhibit Staphylococcus aureus and Candida albicans of all compounds showed that 2 from N. sessilifolia was inhibitory to the growth of Staphylococcus aureus with MIC 25 ug/disc, compounds from L, pinciana. the activity of 10 to inhibit the growth of Staphylococcus aureus with MIC 0.5 mg/ml and MBC 10 mg/ml, and 2 was inhibitory to the Candida albicans with MIC 1 mg/ml and MBC 5 mg/ml. the compounds isolated from three plants were tested the activity to antibiotic resistance Staphylococcus aureus, 10 compounds including one kind of the type of anthraquinones, one compound of the type of aromatic derivative, one of the type of flavane, and seven of the type of triterpene and glycoside. During to observe the result of cooperation the penicillin and Trirnethoprim with those compounds from aboved plants, several compounds had the activity to inhibit antibiotic resistance bacteria strain {Staphylococcus aureus}. compound 21 (MIC 0.57 y. g /disc) from Mitragyna rotundifolia., and 23 (MIC 0.565 u g /disc) from L. pinciana was the similar result to inhibit antibiotic resistance bacteria as that of the amoxillin and clavulanate potassium chewable tablets. On the basis of above-mentioned, the mechanism of inhibition to antibiotic resistance bacteria was discussed by the liquid cultivation, and established the growth curve of CNEC26003 strain to determine the inhibitory ratio of the compound 10 (IC_(50)=7.39 jig/ml 12hrs; 12μg/ml 24hrs), 20 (IC_(50)=14.27μg/ml 12hrs; 50.18μg/ml 24hrs)from L pinciana and 21 (ICS0=45.99μg/ml 12hrs; 58.53 ug/ml 24hrs) from M. rotundifolia Ethanol extract from Neonauclea. sesslifolia Merr., Luculia. pinciana Hook, and Mitragyna. rotundifolia (Roxb.) Kuntze. showed activity on inhibition of P388 mouse leukaemia and A-549 human lung cancer cell line, and compounds isolated from those plants were assayed the activities to inhibition of HL-60 hunman leukaemia and Hela Cervical Cancer Cell line. 19 from Luculia. pinciana Hook., at the concentration of 1, 10 and 100μg/ml, the inhibitory rates were 11.3, 24.7 and 58.0% respectively to the HL-60 leukaemia cell line using the method of MTT. The antibiotics and resistance bacteria were reviewed.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.kib.ac.cn/handle/151853/682
Appears in Collections:昆明植物所硕博研究生毕业学位论文_学位论文

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三种茜草科植物化学成分及其抗菌活性研究.康文艺[d].中国科学院昆明植物研究所,2003.20-25
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