黄河三角洲滨海湿地药用耐盐植物蒙古鸦葱和柽柳化学成分及生物活性研究
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摘要
滨海湿地处于海陆交汇地带,受海陆共同作用,是一个特殊的动植物生长区。为适应特定的生长环境,滨海湿地植物可能具有独特的体内代谢途径并产生特殊的化学防御体系以维持生存,同时也为获得高效低毒的药用生物活性成分提供了新的来源。
     黄河三角洲滨海湿地位于山东省东营市黄河入海口,北临渤海,东靠莱州湾,是世界上暖温带最完整、最广阔、最年轻及生物多样性最为丰富的典型滨海湿地。湿地中动植物资源丰富,其中,药用植物蒙古鸦葱为民间常用中草药,具有清热解毒、消肿散结、消炎、通乳之功效。主治疔疮痈疽、乳痈、跌打损伤、劳伤、乳腺炎和毒蛇咬伤。柽柳为药典收载传统中药,具有有疏风、解表、透疹、解毒之功效。主治风热感冒、麻疹初起、疹出不透、风湿痹痛和皮肤瘙痒。为了寻找高效低毒生物活性成分,我们对黄河三角洲滨海湿地典型的药用耐盐植物蒙古鸦葱Scorzonera mongolica Maxim.和柽柳Tamarix chinensis Lour.进行了系统的化学成分及抗肿瘤生物活性研究:
     (1)采用高温消解,电感耦合等离子体质谱法( ICP - MS)测定了蒙古鸦葱S. mongolica Maxim.和柽柳T. chinensis Lour.中27个无机元素。蒙古鸦葱中Ce、K、Al和Ca含量最高,微量元素Ce、Fe、Mn、Zn和Cu含量较高;而柽柳中K、Na、Ca和Mg含量最高,微量元素Fe、Zn、Cr、Mn和Se含量较高。
     (2)采用气相色谱/质谱法(GC/MS)测定了蒙古鸦葱S. mongolica Maxim.和柽柳T. chinensis Lour.的挥发油成分。鉴定了蒙古鸦葱挥发油中的13个成分,占挥发油总量的93.43%。2种主要成分为三十一烷(34.75%)和何帕-22(29)-烯-3β-醇(21.47%),占挥发油总量的56.22%;检出了柽柳挥发油中的26个成分,占挥发油总量的91.0%。4种主要成分为十六酸甲脂(29.20%)、十八碳二烯酸甲脂(20.88%)、9-十八碳烯酸甲酯(12.31%)和β-维生素E (7.65%),占挥发油总量的70.0%。
     (3)采用氨基酸自动分析仪测定了蒙古鸦葱S. mongolica Maxim.和柽柳T. chinensis Lour.中氨基酸的组成及含量。蒙古鸦葱S. mongolica Maxim.中总氨基酸含量为5.7318%,必需氨基酸含量为1.8600%,中性氨基酸含量为3.1694%,酸性氨基酸含量为1.5640%,碱性氨基酸含量为0.9984%。其中,半胱氨酸、天冬氨酸、精氨酸和谷氨酸含量分别为0.8839%、0.8800%、0.7883%和0.6840%。柽柳T. chinensis Lour.总氨基酸含量为6.3336%,必需氨基酸含量为2.8494%,中性氨基酸含量为4.2250%,酸性氨基酸含量为1.1990%,碱性氨基酸含量为0.9096%。半胱氨酸、谷氨酸、精氨酸和亮氨酸含量分别为0.9457%、0.7197%、0.5866%和0.4958%。
     (4)以细胞毒活性为导向,采用D101大孔树脂分配色谱、正相硅胶色谱、Sephadex LH-20凝胶色谱和制备HPLC等色谱分离手段,运用IR、MS、HRMS、1H-NMR、13C-NMR(DEPT)、1H-1HCOSY、HMQC、HMBC、NOESY等结构鉴定技术,自蒙古鸦葱S. mongolica Maxim.中分离鉴定了23个化合物。其中三萜类成分19个:分别3β-十四酰桑二醇(SM-1)、3β-十二酰桑二醇(SM-2)、羽扇豆醇(SM-3)、白桦脂醇(SM-4)、白桦脂酸(SM-5)、乙酰羽扇豆醇(SM-6)、23Z-3β-acetoxyeupha-7, 23-diene-25-ol(SM-7)、Dammar-20-ene-3β-ol, 24-methy- lene- acetate(SM-8)、3β-齐敦果烷乙酸酯(SM-9)、蒲公英甾醇(SM-10)、蒲公英甾醇乙酸酯(SM-11)、伪蒲公英甾醇(SM-12)、伪蒲公英甾醇乙酸酯(SM-13)、3α-香树脂醇乙酸酯(SM-14)、达玛-24-烯-3β-十四酰氧基-20S-醇(SM-15)、丁酰鲸鱼酸乙酸酯(SM-16)、multiflorenyl acetate(SM-21)、3β-十四酰高根二醇(SM-22)和3β-十二酰高根二醇(SM-23);倍半萜类成分1个:脱氢木香内酯(SM-17);甾醇类成分3个:分别是β-谷甾醇(SM-18)、胆甾醇(SM-19)、5α,8α-环二氧-24-甲基胆甾-6,22-二烯-3β-醇(SM-20)。其中,化合物SM-1、SM-2、SM-22和SM-23为新化合物;SM-4、SM-5、SM-7、SM-8、SM-10、SM-11、SM-12、SM-13、SM-14、SM-15、SM-16、SM-17、SM-20和SM-21首次从该属中分离得到;所有成分均为首次从该种中分离得到。
     (5)从柽柳T. chinensis Lour.中分离得到了27个单体化合物。其中,甾醇类成分6个:分别是β-谷甾醇(TC-1)、豆甾-4-烯-3,6-二酮(TC-2)、麦角甾-4,24(28)-二烯-3–酮(TC-3)、豆甾烷-3,6-二酮(TC-4)、豆甾-4-烯-3-酮(TC-5)、胆甾醇(TC-6);三萜类成分8个:分别是羽扇豆醇(TC-7)、白桦脂醇(TC-8)、myricadiol(TC-9)、isomyricadiol(TC-10)、isoaleuritolic acid(TC-11)、isoaleuritolic acid 3-p-hydroxy cinnamate(TC-12)、2α-羟基齐墩果酸(TC-13)、24-亚甲基环阿尔廷醇(TC-14);二萜类成分1个:植醇(TC-15);黄酮类成分7个:分别是5-羟基-7,4'-二甲氧基黄酮(TC-16)、山奈酚(TC-17)、山奈酚-4′-甲醚(TC-18)、山奈酚-7-甲醚(TC-19)、山柰酚-4′,7-二甲醚(TC-20)、槲皮素(TC-24)和槲皮素-3',4'-二甲醚(TC-25);酚酸类成分5个:邻苯二甲酰二异丁酯(TC-21),邻苯二甲酰正二丁酯(TC-22)、4-羟基苯甲酸(TC-23)、和和阿魏酸(TC-23)。其中,化合物TC-2、TC-3、TC-4、TC-5、TC-11、TC-13、TC-14、TC-16和TC-27为首次从该属植物中分离得到;化合物TC-7、TC-8、TC-9、TC-10、TC-15、TC-21、TC-22、TC-23和TC-26为首次从柽柳T. chinensis Lour.中分离得到。
     (6)采用MTT、SRB法,对分离得到的部分化合物进行了抗肿瘤活性的筛选,化合物SM-4、SM-5、SM-6和SM-7、TC-2、TC-12和TC-13和对人肺癌细胞A-549、TC-15对人肝癌细胞显示有意义的细胞毒活性。
Organisms of coastal wetlands, effected by oceanic and terrestrial environment, likely produce more unique secondary metabolites with various biological activities. At present, a lot of structurally novel and bioactive secondary metabolities, including sesquiterpenes, diterpenes, triterpenes, steroids, ceramides and somon, have been isolated from them. These constituents exhibited potential bioactivities such as antiflammation, antitumor, antifouling and cytotoxic activities. All these will provide more opportunities to find lead compounds.
     Yellow River Delta wetland is the youngest, broadest and the most integrated wetland ecosystem in the warm-temperate zone of the world. It posseses the rich and diverse biological resource. Among them, the family Compositae plant Scorzonera mongolica Maxim. and the family Tamaricaceae plant Tamarix chinensis Lour. were used as Traditional Chinese Medicine(TCM) or folk herbal drug the past time. However, no reports have been found about the chemical research on these plant to date. In order to find bioactive components, we systematically investigated the constituents of S. mongolica Maxim. and T. chinensis Lour. collected from Dongying, Shandong Province of China, including amino acids, organic elements, volatile oils and other organic compounds.
     The MeOH extracts of two species showed significant cytotoxic activities against selected tumor lines, mouse leukemia cell (P388), human hepatic carcinoma cell (BEL-7402), human lung cancer cells (NCI-H460), Breast Carcinoma Cell Line (MCF7), human glioma cell line (SF-268) and Human lung adenocarcinoma epithelial cell line (A-549), respectively. Bioassay-guided isolation resulted in 50 components, 4 new ones as well as 46 known ones. Their structures and stereochemistry were determined on the basis of MS, IR, 1H-NMR, 13C-NMR (DEPT), extensive 2DNMR (1H-1HCOSY, HMQC, HMBC, NOESY) spectroscopic analysis and comparison with the literatures.
     (1) The inorganic element of the sample from S. mongolica Maxim. and T. chinensis Lour. were measured by ICP-MS. 27 elements were detected in the two species. The main inorganic elements were Ce, K, Al, Ca and Fe and the main trace elements were Ce, Ga, Mn, Zn and Cu. in S. mongolica Maxim. respectively. While in T. chinensis Lour., the main inorganic elements were K, Na, Ca and Mg, the main trace elements were Ce, Fe, Mn and Zn.
     (2) The determination of Volatile oils was finished by GC-MS. The results showed that the plant S. mongolica Maxim. was composition of 13 ones, and 2 main components that Hentriacontane (34.75%) and A? - neogammacer - 22(29) - en - 3β- ol(21.47%) account for 93.43% in total Volatile oils. The Volatile oils of T. chinensis Lour. contains 26 components, among them, 4 ones including Hexadecanoic acid methyl ester (29.20%), 11,14-Octadecadienoic acid methyl ester (20.88%), 9-Octadecenoic acid methyl ester (12.31%) andβ-Tocopherol (7.65%) account for 70.0% in total Volatile oils.
     (3) The amino acid analysis was determined on the auto-aminoacid analyzer. 16 kinds of amino (5.7318%) were identified, the main components were Cys(0.8839%), Asp(0.0.88001%) and Arg(0.7883%) in S. mongolica Maxim., respectively. The species T. chinensis Lour. also contained 16 kinds of amino acids(6.3336%), the main components were Glu(0.7197%), Arg(0.5866%) and Cys (0.9457%).
     (4) Guided by cytotoxic activities, 23 Compounds, 3β–teradecanoyloxy-28– hydroxyl–olean–18–ene (SM-1), 3β–dodecanoyl–28–hydroxyl–olean–18–ene (SM-2), lupeol (SM-3), betulin (SM-4), betulinic acid (SM-5), lupeo acetate (SM-6), 23Z–3β–acetoxyeupha–7, 23–diene–25–ol (SM-7), Dammar–20-ene-3β-ol, 24– methylene–acetate (SM-8),β–pulcherryl acetate (SM-9), taraxasterol (SM-10), Taraxasterol acetate (SM-11),ψ–taraxasterol (SM-12),ψ–taraxasteryl acetate (SM-13),α–Amyrin acetate (SM-14), Dammar–24–ene-3, 20–diol (SM-15), 3β–tetradecanoate, butyrospermol acetate (SM-16), Dehydrocostus lactone (SM-17),β-sitosterol (SM-18), cholesterol (SM-19), 5α. 8α-epidioxy-(22E, 24R)-stigmaster- 6, 22- dien-3β-ol (SM-20), multiflorenyl acetate (SM-21), 3β–teradecanoyloxy-28–hydroxyl–olean–12–ene (SM-22) and 3β– dodecanoyl–28–hydroxyl–olean–12–ene (SM-23),were obtained from S. mongolica Maxim. Among them, SM-1, SM-2, SM-22 and SM-23 were new triterpenes; SM-4, SM-5, SM-7, SM-8, SM-10, SM-11, SM-12, SM-13, SM-14, SM-15, SM-16, SM-17, SM-20 and SM-21 were isolated from the genus Scorzonera for the first time. All components were isolated firstly from the species S. mongolica Maxim.
     (5) 27 Compounds were obtained from T. chinensis Lour. Based on spectroscopic analysis and comparison with the literatures, their sructures were identified asβ–sitosterol (TC-1), stigmast–4–ene-3, 6–dione (TC-2), Ergosta–4, 24(28)–diene–3–one (TC-3), stigmast–4–ene-3, 6-dione (TC-4), sitoste rone (TC-5), cholesterol (TC-6), lupeol (TC-7), betulin (TC-8), myricadiol (TC-9), isomyricadiol (TC-10), isoaleuritolic acid (TC-11), isoaleuritolic acid 3-p-hydroxy cinnamate (TC-12), maslinic acid (TC-13), 24–methylenecycoartanol (TC-14), Phyto (TC-15), 5-hydroxy–7, 4?–dime–thoxyflavone (TC-16), Kaempferol (TC-17), 4′- methylkaempfero (TC-18), 7-methylkaempfero (TC-19), 4′,7–dimethyl– kaempfero (TC-20), diiso–butyl–O–phthalate (TC-21), dibutyl phthalate (TC-22), 4-Hydroxy -benzoic acid (TC-23), Quercetin (TC-24), 3',4'-dimethylquercetin (TC-25), ferulic acid (TC-26) and methyl-P-hydrobenzoate (TC-27). Among them, TC-2, TC-3, TC-4, TC-5, TC-11, TC-13, TC-14, TC-16 and TC-27 were isolated from the genus Tamarix for the first time. TC-7、TC-8, TC-9, TC-10, TC-15, TC-21, TC-22, TC-23 and TC-26 were isolated firstly from T. chinensis Lour.
     (6) The anti-tumor activities of 23 compounds were tested by MTT and SRB methods. SM-4、SM-5、SM-6、SM-7、TC-2、TC-12、TC-13and TC-15 showed potential anti-tumor activities.
引文
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