麻叶荨麻及活血益气汤活性部位的化学成分研究

【作者】 马学敏；

【导师】 郭亚健；

【作者基本信息】 北京中医药大学 ， 中药化学， 2004， 博士

【摘要】 荨麻属植物(Urtica)麻叶荨麻(Urtica cannabina L.)，在我国药用历史悠久，西北地区民间常用于治疗风湿关节炎、荨麻疹、蛇咬伤、小便不利等症。具有祛风除湿，活血解痉，抗炎消肿、利尿等功效。麻叶荨麻的同属植物异株荨麻在欧洲同样有悠久的药用历史，国外现阶段已开发出多种保健品及治疗风湿关节炎、良性前列腺增生(BPH)的药物。在国内研究已相对滞后的情况下，为充分利用西北地区丰富的植物资源，我们对麻叶荨麻的化学成分进行了比较系统的研究， 以期为进一步的研究开发奠定基础。 采用色谱技术对麻叶荨麻的乙醇提取物进行分离，得到了13个化合物。通过化学方法和光谱技术鉴定了其中10个化合物，分别为两个香豆素类化合物：6，7-二羟基香豆素(Ⅰ)，7-羟基-6—甲氧基香豆素(Ⅱ)；两个香豆素类的前体化合物：对羟基桂皮酸(Ⅲ)、对羟基桂皮酸甲酯(Ⅳ)；两个烷烃类化合物：正三十一烷(Ⅴ)正二十九烷(Ⅵ)；另外还；有3，5-二甲氧基-4-羟基苯甲醛(Ⅶ)，山奈酚7-0-鼠李糖苷(Ⅷ)， β-谷甾醇(Ⅸ)，胡萝卜苷(Ⅹ)。其中，除化合物Ⅸ、Ⅹ外，其余均首次从该植物中分离得到，化合物Ⅲ—Ⅷ，为首次从荨麻属植物中得到。 对分离过程中得到的脂肪油，通过衍生化后将其制成甲酯化物。利用GC-MS技术，从NIST谱库中检索到62个色谱峰，鉴定了其中25个脂肪酸，占出峰面积的74.04％。其中棕榈酸含量最高，达23.23％。其余含量较高的还有硬脂酸、花生酸、二十一烷酸、山嵛酸、二十四烷酸。 有机酸成分是荨麻是荨麻属植物主要抗炎活性成分之一。为研究麻叶荨麻中的有机酸成分，我们按有机酸类成分的提取方法提取总有机酸，进行甲酯化后，利用GC-MS技术从NIST谱库中共检索到60个色谱峰，鉴定了其中27个峰，占总出峰面积的71.63％。其中对羟基桂皮酸含量最大，占27.86％。其余含量超过3.00％的还有顺式乌头酸，对羟基苯甲酸，香草酸，2，5-二羟基苯甲酸，壬二酸，柠檬酸。已知对羟基桂皮酸、顺式乌头酸、对羟基苯甲酸、香草酸、柠檬酸具有抗炎活性。 肝纤维化是各种慢性肝病发展至肝硬化的一个必经共同途径，阻止肝纤维化麻叶尊麻及活血益气汤活性部位的化学成分研究的形成对防止肝硬化具有重要意义。活血益气方药是王健等人根据中医理论及临床经验配伍组成的抗肝纤维化方药，由黄茂，大黄，何首乌，丹参，桃仁五味中药组成。药效学试验表明具有较好的抗猪血清诱导的大鼠实验性肝纤维化作用，其有效部位有两种不同的作用模式:一种是以水溶性成分为主的早期生效、具有一定程度的剂量依赖性抑制作用、抑制效应较弱的效应模式;另一种是以脂溶性成分为主的生效较晚、有明显的时间依赖性抑制作用、抑制效应较强的效应模式。 其中脂溶性部位Alc950/0和Alc5006在很低的剂量下，仍显示出强烈的抑制效应为阐明活性作用的物质基础，我们对Alc95%和Alc50%的化学成分进行了研究。经过溶剂萃取、大孔吸附树脂富集、sephadex LH一20精制、制备TLC及Semi一pre HpLC等手段从中分离了20个化合物，利用理化性质和波谱技术(uv、工R、El/MS、Esl/MS、，H一NMR、”C一NMR、oEPT、HMQe、HMBC)相结合鉴定了创门的结构，分别为9种葱醒类化合物:大黄酚(Chrys。phanol)，大黄素甲醚(physeion)，大黄素(emodin)，芦荟大黄素(aloemodin)，大黄酸(rhein)，番泻普A或B(sennoside Ao:B)，。一经基大黄素(。一hydroxylemodin)，大黄酚一8一0一p一D一葡萄糖普(ehrys。phan。l一8一。一口一D一glueoside)，芦荟大黄素一。一o一p一D一葡萄糖普(aloemodin一。一0一p一D一glueoside):3种菲醒类化合物:丹参酮IIA(tanshinoneIIA)，隐丹参酮(e ryptotanshinone)，丹参酮I(tanshinonel):1种异黄酮:7，3‘一二经基一5‘一甲氧基异黄酮(7，3‘一dihydroxy一5‘methoxyisofla-vone)以及原儿茶醛(protoeaeehuic aldehyde)，原儿茶酸(pr。t。eaeehuicaeid)，黄茂甲普(astragal。side Iv)，苦杏仁普(amygdazin)，z，3，s，4‘一四经基二苯乙烯一2一o一口一D一葡萄糖普(z，3，s，4‘一tetrahydr。xylstilbene一2一。一口一D一glueoside)，(+)一儿茶素[(+)一catechin]，桂皮酸(einnamse acid)。其中7，3’一二经基一5‘一甲氧基异黄酮是自然界中罕见的B环为间位二取代的异黄酮类化合物，为新化合物。 采用高效液相色谱法，5 p moiamonsil一C，:分析柱，甲醇一0.1%磷酸(87:13)洗脱，436nm检测，同时测定了Alc95%部位中5个游离葱醒的含量，结果芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的平均回收率分别为98.30%、97.12%、98.18%、97.66%、97.16%、RSD分别为1.64%、1.15%、0.95%、1.51%、1.64%(n二5)o方法操作简便、准确、重现性好，可用于该活性部位的质量分析。更多还原

【Abstract】 Studies on the chemical constituents of Urtica Cannabina L. and the active parts of the Prescription for benefited Qi to Promote flow of blood.Urtica Cannabina L. is one species of Urtica. There are very rich plant resources in the northwest of China. The local people use U. cannabina to care rheumatism, nettle rash, wounds bited by snakes and so on. It was said to have the function of driving away wind-dampness, opening channels and collaterals, relaxing convulsions, antiinflammation and detumescence. While, Urtica dioica is another species of Urtica as same as U. Cannabina .It widely distributed in the Europe and had been used for a long time. From the 1960’s to now, the foreign pharmaceutists had developed a lot of healthness protecting products and OTC drugs. Which are used for the therapy of rheumatism and benign prostate hyperplasia (BPH). In order to make full use of the plant resource in the northwest of China and the information source of foreign countries, we carried on a system research on the chemical constituents of U. Cannabina L.By using different chromatography techniques, 13 compounds were isolated from the 80% EtOH extract. Based on modern spectroscopic analysis technique and chemical methods, the structures of 10 compounds were elucidated as esculetin, scopoletin, p-coumaric acid, methyl P-coumarate, hentriantane, nonacosane, 3,5-dimeth--oxy-4-hydroxybenzaldehyde,Kaempfero1-7-0-rhamnoside, -sitosterols and daucosterol.This is the first report that compounds are in Urtica genus and all compounds except IX and X were first reported in Urtica cannabina L.Fatty acid from petroleum part was subjected to GC-MS after saponificated and methanolized. The derivative were composed of 62 components, 25 of which were identified, account for 74.04%. The content of palmitic acid was 23.23%, while stearic acid, eicosanoic acid, heneicosanoic acid, docosanic acid and tetracosanoic acid were more than 3% respectively.Organic acid was one of anti-inflammation active component in Urtica plant. General organic acid was extracted and methanolized to be analyzed by GC-MS. 60 peaks were inspected in NIST Dictionary, 27 peaks wereidentified, account for 71. 63%. The content of p-coumaric acid was 27. 86%, while cis-aconitic acid, methyl paraben, vanillic acid, 2,5-dihydroxy-benzoic acid, nonane dioic acid, citric acid were more than 3% respectively. Among them, p-coumaric acid, cis-aconitic acid, vanill ic acid, citric acid were well known antiinflammation active components. Fumaric acid, citric acid, succinic acid existed in Urtica Cannabina L. as well as in Urtica diocio.The severe consequence of hepatic fibrosis is cirrhosis liver . Preventing hepatic fibrosis is an important target to treat cirrhosis of liver. The preseription for benefited Qi to promote flow of blood was made up by Jian Wang et al. based on Chinese Traditional Medical Principle and clinical experience, which concludes Rheum palmatum L., Salvia miltinrrhiza Bge, Astragalus membranaceus (Fisch) Bge .var. mongholicus (Bge),Hsiao, Prunus PersicacL. Batsch, Polygonum multiflorum Thunb. It shows protective effects against rat Hepatic fibrosis in vivo. Lipophilic extracts show significant inhibiting effect at very low concentration.To further study the chemical constitutes of lipophilic active parts , we prepared Alc95% and Alc50% according to original literature. 20 compounds were isolated by using macroporous resin, silica gel, polyamide, Sephadex LH-20, Rp-18, preparative TLC and preparative HPLC chromato--graphy. The structures of them were elucidated by means of physical and chemical properties, modern spectroscopic analysis techniques桿V, IR, EI/MS,ESI/MS,1H-NMR,13C-NMR,DEPT,HMQC,HMBC. They are nine anthraquinones: chrysophanol (95-1), physcion (95-2), emodin (95-5), aloemodin (95-7), rhein (50-1) , sennoside A or B(50-6) , -hydroxylemodin (95-10) , chrysophano1-8-0-D-glucoside (95-12), aloemodin-0-D-glucoside (95-13); three phenanthaquinones: tanshinone A (95-3), cryptotanshinone (95-6) , tanshinone I (95-4); one isoflavone: 7,3’ -dih更多还原