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题名: 中亚沙棘化学成分与保肝有效组分研究
作者: 李寅庆
答辩日期: 2014-05-30
导师: 阿吉艾克拜尔·艾萨
专业: 有机化学
授予单位: 中国科学院大学
授予地点: 北京
学位: 博士
关键词: 中亚沙棘 ; 化学成分 ; 黄酮 ; 油脂 ; 总三萜 ; 总酚 ; 保肝
摘要: 沙棘是一种世界范围内分布十分广泛的植物,《中国药典》中记载其有健脾消食,止咳祛痰,活血散瘀的功效。中亚沙棘(Hippophae rhamnoides L. subsp. turkestanica)是胡颓子科(Elaeagnaceae)沙棘属(Hippophae L.)鼠李沙棘(Hippophae rhamnoides)的亚种,在我国仅生长于新疆地区,维吾尔药中称之为“吉汉”,《维吾尔药志》中记载其果实主治湿热性或血液质性疾病,如热性咳嗽,胃热纳差,恶心呕吐,顽痰不化,小便不利,肝硬腹水,高血压病,气喘心悸。 近年来,全世界范围内沙棘资源的研究与开发有了长足进展,与沙棘相关的产品层出不穷。而对于新疆特有的中亚沙棘相关研究报道寥寥无几。中亚沙棘含有哪些特别的化学成分?中亚沙棘在哪些主要的生物活性成分含量上相比其他品种沙棘具有显著的优势?这些问题依然模糊不清,而中亚沙棘绝大多数相关的产品开发也仅限于简单的食品加工。本论文以新疆特有的中亚沙棘果实为主要的研究对象,对其化学成分进行系统深入的研究,对其保肝有效组分提取纯化工艺进行优化,并对有效组分的保肝活性进行评价。本论文第一章对沙棘属植物的分类及资源地理分布和近年来沙棘属植物化学成分的研究及生物活性的研究进行了系统的阐述,并对目前沙棘资源的开发现状进行了简单的介绍。第二章展开对中亚沙棘化学成分的基础研究,从正丁醇部位和乙酸乙酯部位中共分离鉴定了21个单体化合物,14个化合物首次从本植物中分离得到,其中的6个化合物首次从胡颓子科植物中分离得到。在此基础上将中亚沙棘与新疆及国内分布较广的四种沙棘中的黄酮苷,酸水解为槲皮素、异鼠李素、山奈酚三种黄酮苷元,并通过HPLC进行含量比较。结果表明:新疆阿克苏乌什县产的中亚沙棘总黄酮苷元含量最高,每克干果中含2.334 mg总黄酮苷元;几种沙棘中含量最高的黄酮苷元均为异鼠李素。对中大极性化学成分进行系统研究后,通过GC-MS的方法对中亚沙棘的油脂成分进行研究。首先比较了中亚沙棘果实中果皮和种子两个部位油脂中脂肪酸成分和非皂化脂质组成的差异。果皮油中棕榈油酸含量最高达总脂肪酸的41%,其非皂化脂质中的角鲨烯和维生素E相比显著高于种油;种油中不饱和脂肪酸含量较高可达总脂肪酸的86%,非皂化脂质中甾醇相比果油含量更高。在此基础上,将中亚沙棘与新疆及国内分布较广的四种沙棘含油率和脂肪酸组成进行了比较。结果表明和田策勒县的蒙古沙棘和阿勒泰清河县的大果沙棘出油率最高达28%,喀什疏附和阿克苏乌什产的中亚沙棘次之。和田策勒产的蒙古沙棘不饱和脂肪酸占70.7%,高于其他品种,乌什产的中亚沙棘达69.5%仅次之,但乌什产的中亚沙棘棕榈油酸含量占总脂肪酸的40.4%显著高于其他样品。第三章在第二章化学成分研究的基础上,结合中亚沙棘在传统维吾尔医中的应用,对中亚沙棘果实总三萜和总酚进行同步提取,并对提取纯化工艺进行优化,最后通过CCl4致大鼠急性肝损伤模型,对其保肝作用进行评价。适宜的提取纯化工艺条件为:90℃下10倍重量的50%乙醇提取3次,每次提取3h,提取液除去乙醇后稀释至0.067 g生药/mL,调节pH至6,上AB-8大孔树脂柱,上样液流速为2 BV/h,上样量为9-10 BV,洗脱剂为70%乙醇,洗脱剂用量为3.5 BV。纯化后每克有效组分含0.49 g齐墩果酸当量的总三萜和0.36 g没食子酸当量的总酚。所得有效组分具有较强的抗ABTS氧化自由基的能力,能明显抑制四氯化碳导致的肝损伤大鼠血清中ALT、AST、TC、TG的升高,中剂量和高剂量的有效组分具有显著的抑制肝细胞坏死和水肿的作用。所得有效组分在新型的沙棘保肝药品及保健品的开发中具有一定的应用前景。
英文摘要:
Hippophae is widely grown throughout the world. Pharmacopoeia of the People’s Republic of China recorded it had a good effect in promoting digestion, relieving cough, eliminating phlegm and improving circulation. As a subspecies of Hippophae rhamnoides, Hippophae rhamnoides L. subsp. turkestanica is exclusively distributed in Xinjiang in our country. Its Uygur medicine name is “Ji Han”. "Uygur medicine Zhi" recorded it had therapeutic value in the treatment of wetness-heat and blood diseases such as cough, stomach anorexia, nausea, vomiting, asthma, much sputum, difficulty in micturition, cirrhosis and hypertension. In recent years, global research and development of the Hippophae resources had made great progress, and a variety of Hippophae products were put on the market. However, the research reports about the Hippophae rhamnoides L. subsp. turkestanica, unique in Xinjiang were very few. What particular compounds are contained in the Hippophae rhamnoides L. subsp. turkestanica? Which kind of bioactive ingredients in the Hippophae rhamnoides L. subsp. turkestanica is more abundant than in other species? These questions were still unclear. The great majority of the Hippophae rhamnoides L. subsp. turkestanica products were confined to simple food processing. In this research, we carried out a systematic study on the chemical composition of Hippophae rhamnoides L. subsp. turkestanica fruits, optimized the extraction and purification process of its hepatoprotective fraction, and evaluated the hepatoprotective activity of the effective fraction. In the first chapter, species of the Hippophae genus and their distribution were introduced. Progress in studies of the Hippophae genus, including chemical composition, biological activity and products development were reviewed here. All of our research on the chemical composition of the Hippophae rhamnoides L. subsp. turkestanica is displayed in the second chapter. 21 compounds were separated and identificated from the n-BuOH and EtOAC fractions. In these compounds, 14 of them were obtained from the Hippophae rhamnoides L. subsp. turkestanica for the first time, and 6 of them were isolated from the Elaeagnaceae family for the first time. The total flavonoid aglycone contents of the Hippophae rhamnoides L. subsp. turkestanica were compared with other main species distributed in China, especially in Xinjiang province by HPLC. Hippophae rhamnoides L. subsp. turkestanica grown in Wushi of Xinjiang had the highest level, each gram of dried fruit containing 2.334 mg total flavonoid aglycone. Isorhamnetin is the highest concentrations flavonoid aglycone in all tested species of Hippophae. After the research on the middle and large polar chemical constituents, we continued to study on the lipid of the Hippophae rhamnoides L. subsp. turkestanica by GC-MS. At first, we compared the fatty acid and non-saponification lipid composition between the seeds and the peel of the Hippophae rhamnoides L. subsp. turkestanica fruits. More palmitoleic acid, squalene and vitamin E were in the peel oil, but more unsaturated fatty acids in the seeds oil. 41% of fatty acid in peel oil was palmitoleic acid, and 86% of fatty acid in seeds oil was unsaturated fatty acids. Then, we compared the fatty acid compostion of the Hippophae rhamnoides L. subsp. turkestanica with other main species which were distributed in China, especially in Xinjiang province. The Hippophae rhamnoides L. subsp. mongolica grown in Cele and the big fruit seabuckthorn distributed in Qinghe had the highest oil yield up to 28%. The Hippophae rhamnoides L. subsp. mongolica grown in Cele contained the most unsaturated fatty acids. 70.7% of fatty acid in it was unsaturated fatty acids. However, the Hippophae rhamnoides L. subsp. turkestanica contained highest level palmitoleic acid. 40.4% of fatty acid in it was palmitoleic acid. In chapter 3, the isolation and purification craft of the hepatoprotective fraction from Hippophae rhamnoides L. subsp. turkestanica was established, based on the chemical composition study in chapter 2 and the traditional Uighur medicine usage. Then, its hepatoprotective effect was evaluated by the model of carbon tetrachloride induced liver injury in rats. The optimal conditions for separation and purification was extract 5 times by 5 BV 50% ethanol in 90℃, 3 hours each time. Then, evaporated to dryness and diluted to 0.067 g crude drug / mL, adjusted pH to 6, chosed the AB-8 macroporous resin column. The flow rate of the sample solution is 2BV / h, and the sample solution volume was 9-10 BV. 70% ethanol was used to eluent, and the eluent volume was 3.5 BV. Each gram final product contained 0.49 OLE g total triterpenoids and 0.36 GAE g total phenolic. The hepatoprotective fraction had a strong ability of inhibition ABTS radical oxidation. It could inhibit the increase of AST, ALT, TG and TC in the blood of liver injurying rats inducting by carbon tetrachloride. Then, middle dose and high dose of the fraction could significantly reduce the necrosis and edema of the liver cells. Therefore, the Hippophae rhamnoides L. subsp. turkestanic hepatoprotective fraction have a potential price in the development of hepatoprotective new drugs and health products.
内容类型: 学位论文
URI标识: http://ir.xjipc.cas.cn/handle/365002/3428
Appears in Collections:资源化学研究室_学位论文

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作者单位: 中国科学院新疆理化技术研究所

Recommended Citation:
李寅庆. 中亚沙棘化学成分与保肝有效组分研究[D]. 北京. 中国科学院大学. 2014.
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