三叶木通化学成分及生物活性研究

发布时间：2018-05-07 19:51

本文选题：三叶木通 + 化学成分　；参考：《陕西科技大学》2017年硕士论文

【摘要】：三叶木通Akebia trifoliate(Thunb.)Koidz为木通科木通属藤本植物,是2015版《中国药典》收载品种木通的基源植物之一,干燥藤茎入药,具有利尿通淋、通经下乳、清心除烦、抗氧化防衰老等功效。临床上以其良好的利尿作用,广泛的用于治疗淋症、心烦尿赤、水肿、湿热痹痛等。然而其发挥利尿作用的活性部位及成分尚不明确。本课题以天然产物化学结合药理实验的方法,拟寻找三叶木通的利尿活性部位及成分,同时对其抗氧化成分进行了研究。三叶木通药材粗粉(4.9 kg),以95%乙醇常温浸提6次,每次浸泡48小时,合并提取液,减压浓缩得到提取物浸膏280.6 g。将浸膏悬浮于水中依次用石油醚、乙酸乙酯、正丁醇进行萃取,减压回收溶剂。得到石油醚萃取物36.8 g(13.1%),乙酸乙酯萃取物29.7 g(10.6%),正丁醇萃取物137.7 g(49.1%),以及剩余水溶性部分70.6 g(25.2%)。采用大鼠代谢笼法对三叶木通藤茎提取物的石油醚、乙酸乙酯、正丁醇萃取物和剩余水溶性部分的利尿活性进行测试。采用硅胶柱色谱及凝胶柱色谱等方法,对利尿作用显著的正丁醇部位(P0.01)进行分离纯化,共得到8个化合物。通过NMR、HRMS等方法,结合文献数据鉴定化合物结构,分别为常春藤皂苷元(1)、丁香酸-4-O-α-L-鼠李吡喃糖苷(2)、β-谷甾醇-3-O-β-D-葡萄糖苷(3)、香草酸葡萄糖苷(4)、丁香酸葡萄糖苷(5)、甘露醇(6)、常春藤皂苷元-3-O-α-L-阿拉伯吡喃糖苷(7)、肌醇(8)。其中化合物2、4、5为首次从木通属植物中分离得到。实验进一步对所分离得到的各化合物进行利尿活性测试,结果表明:与生理盐水对照组相比,甘露醇(P0.05)、常春藤皂苷元-3-O-α-L-阿拉伯吡喃糖苷(P0.05)及常春藤皂苷元(P0.05)的利尿活性显著。采用DPPH自由基清除法,首次对三叶木通乙醇提取物的石油醚、乙酸乙酯、正丁醇萃取物以及水溶性部分进行抗氧化活性筛选。采用多种色谱技术,对抗氧化活性较强的乙酸乙酯萃取物进行化合物分离,共得到9个化合物。根据各化合物的理化性质及核磁数据等解析其结构,分别为β-谷甾醇(9)、香草酸(10)、6,7,10-三羟基-8-十八烯酸(11)、齐墩果酸(12)、2,6-二甲氧基~(-1),4-苯醌(13)、三十四烷(14)、秦皮乙素(15)、咖啡酸(16)、木通苯乙醇苷B(17)。其中化合物10、13、14为首次从木通属植物中分离得到。对各化合物的抗氧化活性进行测定,结果表明:咖啡酸(IC50=14.939μg.mL~(-1))和秦皮乙素(IC50=15.631μg.mL~(-1))对DPPH自由基的清除能力较强,与阳性对照抗坏血酸(IC50=17.269μg.mL~(-1))相当,是三叶木通具有抗氧化作用的物质基础。
[Abstract]:Akebia trifoliata (Akebia trifoliate(Thunb.)Koidz) is one of the basic plants in the 2015 edition of Chinese Pharmacopoeia, which belongs to Liana. The dried vine stems are used in medicine, which have the functions of diuretic entrainment, menstrual circulation, antioxidation, anti-aging and so on. Clinical with its good diuretic effect, widely used in the treatment of leprosy, irritation, edema, damp-heat arthralgia and so on. However, the active sites and components of diuretic action are unclear. The aim of this study was to find out the diuretic active parts and components of Akebia trifoliata by chemical and pharmacological experiments of natural products, and to study the antioxidant components of Akebia trifoliata. The crude powder of Akebia trifoliata was extracted with 95% ethanol at room temperature for 6 times, each time for 48 hours, then the extract was combined with the extract, and the extract was extracted by decompression and concentration, and the extract was 280.6 g. The extract was extracted by petroleum ether, ethyl acetate and n-butanol, and the solvent was recovered by decompression. The petroleum ether extract (36.8 g / L), ethyl acetate extract (29.7 g / L), butanol extract (137.7 g / L) and residual water-soluble fraction (70.6 g / L) were obtained. The diuretic activities of petroleum ether, ethyl acetate, n-butanol extract and residual water-soluble fraction of the extracts from the stems of Akebia trifoliata were determined by the metabolic cage method in rats. Eight compounds were isolated and purified by silica gel column chromatography and gel column chromatography. The structures of the compounds were identified by means of NMR-HRMS and literature data. They are Ivy saponins (1), eugenic acid (-4-O- 伪 -L- rhamopyranoside), 尾 -sitosterol -3-O- 尾 -D- glucoside (3-O- 尾 -glucoside), glucocorticoside (4), riboside (5), mannitol (6), ivy saponins (3-O- 伪 -L-arabinopyranoside) (7N), inositol (8m). Among them, compound 2o 4N 5 was isolated from Akebia for the first time. The diuretic activity of the isolated compounds was further tested. The results showed that the diuretic activities of mannitol P0.05, ivy saponin -3-O- 伪 -L-arabyranoside (P0.05) and Ivy saponin (P0.05) were significant compared with the control group. The antioxidant activity of petroleum ether, ethyl acetate, n-butanol extract and water-soluble part of ethanol extract of Akebia trifoliata was screened by DPPH radical scavenging method for the first time. Nine compounds were isolated from ethyl acetate extract with strong antioxidant activity by various chromatographic techniques. According to the physicochemical properties and nuclear magnetic data of the compounds, The results were as follows: 尾 -sitosterol (9), vanillic acid (10), trihydroxy-8-octadecanoic acid (11), oleanolic acid (Oleanolic acid) (12), dimethoxy (6-dimethoxy) -4-benzoquinone (13), 34 alkane (14), palmatine 1515, caffeic acid (16), xylotene ethanoside (B _ (17). Compound 10C 13N 14 was isolated from Akebia for the first time. The antioxidant activity of each compound was determined. The results showed that the scavenging ability of caffeic acid IC50 (14.939 渭 g 路mL 路L ~ (-1) and cinnamonine (IC50 ~ (15. 631) 渭 g 路mL ~ (-1) on DPPH free radical was stronger than that of the positive control group (IC5050 ~ (17.269 渭 g 路mL ~ (-1), which was the material basis of antioxidation of Akebia trifoliata.
【学位授予单位】：陕西科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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